JCSS级别阴离子标准溶液2


产品编号 产品名称 产品规格 产品等级 产品价格
010-26361 Anion Mixture Standard Solution 2
阴离子混合标准溶液2
50mL JCSS

JCSS级别阴离子标准溶液2JCSS级别阴离子标准溶液2

 

  近年来,随着水道法的修正,可溯源的标准品、标准溶液的使用日渐增多。

  最近公布的《健水发0330水质基准第1号(2016年3月30日附),厚生劳动大臣通过的水质相关的省令规定基准方法等一部分修正内容的注意事项》中,厚生劳动大臣通过的以水质基准相关的省令规定基准方法(2013年厚生劳动省告示第261号),添加了国家计量标准中心认证的可溯源标准原液,同时确定了一定条件下可保证追溯性的标准溶液和混合标准溶液(附带计量法基准证书)。


 ◆阴离子标准溶液2


  本品是水道法水质基准中已被设定基准值的“硝酸盐”“亚硝酸盐”“氟”“氯离子”的混合标准溶液。

  根据厚生劳动大臣通过的以水质基准相关省令规定基准方法《别表第13 离子色谱(阴离子)同时分析法》记载的混合标准溶液比率配制。

 

◆标准原液和JCSS阴离子混合标准溶液2 对比表


水质基准

基准值

(mg/L)

标准原液(别表第13)

(mg/mL)

混合标准溶液(别表第13)

(mg/mL)

JCSS成分

(产品编号:010-26361)

浓度

(mg/L)

硝酸盐和亚硝酸盐

10以下

1

(硝酸盐)

0.002

(2mg/L)

硝酸盐

20

亚硝酸盐

0.04以下

1

0.001

(1mg/L)

亚硝酸盐

10

0.8以下

1

0.005

(5mg/L)

氟化物

50

氯离子

200以下

1

0.02

(20mg/L)

氯离子

200

 


外泌体分离用单克隆抗体 Anti CD9,CD63,CD81


产品编号 产品名称 产品规格 产品等级 产品价格
CAC-SHI-EXO-M01-50UL Anti CD9 for Exosome Isolation
CD9 外泌体提取抗体
50μL
CAC-SHI-EXO-M01-100UL Anti CD9 for Exosome Isolation
CD9 外泌体提取抗体
100μL
CAC-SHI-EXO-M02-50UL  Anti CD63 for Exosome Isolation
  CD63 外泌体提取抗体 
50μL
CAC-SHI-EXO-M02-100UL  Anti CD63 for Exosome Isolation
外泌体分离抗体CD63 
100μL
CAC-SHI-EXO-M03-50UL  Anti CD81 for Exosome Isolation
  CD81 外泌体提取抗体 
50μL
CAC-SHI-EXO-M03-100UL  Anti CD81 for Exosome Isolation
  外泌体分离抗体CD81 
100μL
CSR-SHI-EXO-K010 ExoTrap Exosome Isolation Spin Column Kit, for Protein Research
外泌体分离亲和柱套装,蛋白研究用
10PREP

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81外泌体分离用单克隆抗体 Anti CD9,CD63,CD81



  本产品是特异性识别外泌体marker CD9,CD63,CD81的抗体,运用免疫沉淀法,分离血清、培养上清液的外泌体。*已获得专利,号码为:PCT/JP2012/083612

背景

  外泌体是由细胞磷脂双分子层分泌形成的细胞衍生囊泡,直径为40nm~100nm。能在活体的唾液、血液、尿液、羊水、恶性腹水等体液中观察得到,也可通过培养细胞分泌得到。近年报道指出,外泌体含有多种蛋白质和RNA,有可能在细胞间的信号传递发挥作用。

 


特点

  ● 高特异性识别外泌体膜蛋白CD9,CD63,CD81。

  ● 1 μg的抗体能100%分离150 μL样品的外泌体。

  ● 对应样品(使用人样品验证)
    CD9:血清、血浆、培养上清液、尿液
    CD63:血清、血浆、培养上清液、尿液
    CD81:血清、血浆、培养上清液

  ● 用于外泌体表面抗原蛋白、内源RNA(miRNA)、蛋白质分析。

应用实例

  1、用Anti-CD9抗体(12A12)进行血清中外泌体的IP-WB

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81


  2、用Anti-CD63(8A12)进行血清中外泌体的IP-WB

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

  3、用Anti- CD81(12C4)进行血清中外泌体的IP-WB

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

  产品说明书请看相关资料

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

外泌体分离用单克隆抗体CD9 CD63 CD81-1611CBAU02.pdf

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81Anti CD9 [ Clone  12A12]

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81Anti CD63 [ Clone  8A12]

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81Anti CD81 [ Clone  12C4]

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81

外泌体分离用单克隆抗体 Anti CD9,CD63,CD81ExoTrapTM Exosome Isolation Spin Column Kit for protein research

参考文献

Nao Nishida-Aoki, Naoomi Tominaga, Fumitaka Takeshita etl. Disruption of Circulating Extracellular Vesicles as a Novel Therapeutic Strategy against Cancer Metastasis[J]Molecular Therapy, 2017, 1, 4, 25(1): 181–191 摘要

                         

内切纤维素酶检测试剂盒

内切纤维素酶检测试剂盒

英文名:Cellulase Assay Kit (CELLG3 Method)

货号:K-CELLG3

规格:180 / 360 assays per kit / 720 (auto-analyser)

市场价: 5537

 纤维素酶是一种重要的酶产品,是一种复合酶,主要由外切β-葡聚糖酶、内切β-葡聚糖酶和β-葡萄糖苷酶等组成,还有很高活力的木聚糖酶。

提供内切纤维素酶检测试剂盒,色度法(400 nm)测定酶制品和发酵产品中的纤维素酶(内切-1,4-β-葡聚糖酶)。

内切纤维素酶检测试剂盒(CELLG3方法) K-CELLG3

使用高纯度β-葡萄糖苷酶和苯亚甲基阻断,2 – 氯-4 – 硝基苯基-β-Dcellotrioside(BClPNPβ-G3)。 β-葡萄糖苷酶的能力确保测定的可靠的最大的灵敏度。BClPNPβ-G3的水解为苯亚甲基,通过纤维素酶阻断纤维二糖和2-氯-4 -硝基苯基-β-D-葡萄糖, 2-Cl-4-硝基苯基-β-D-葡萄糖通过β-葡萄糖苷酶立即裂解为D-葡萄糖和游离的2 – 氯-4 – 硝基苯酚(ClPNP)。

反应时间:16min

适用样品:酶制品和发酵产品

优点:

价格低廉(每次检测成本)

特异性高

方法简单

包含标准品

 

The CELLG3 assay reagent for the measurement of endo-cellulase (endo-1,4-β-glucanase) contains two components;

1) 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-D-cellotrioside (BCNPG3) and 2) thermostable β-glucosidase. The benzylidene blocking group prevents any hydrolytic action by the β-glucosidase on BCNPG3.  Incubation with an endo-cellulase generates a non-blocked colourimetric oligosaccharide that is rapidly hydrolysed by the ancillary β-glucosidase.  The rate of formation of 2-chloro-4-nitrophenol is therefore directly related to the hydrolysis of BCNPG3 by the endo-cellulase.  The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH 9.0).
Please note that a new assay kit (K-CELLG5) is now available for the measurement of endo-cellulase.  The CELLG5 reagent contains a cellopentaose core and exhibits vastly improved sensitivity for some cellulases.  In addition, the exchange of the benzylidene blocking group in CELLG3 for 3-keto-butylidene in CELLG5 improves the substrate’s water solubility significantly, allowing for a reduction in the concentration of DMSO required in the assay.  As DMSO is known to inhibit certain cellulases, this is another benefit in using CELLG5.  Megazyme now recommends the use of K-CELLG5 for all assays for the measurement of endo-cellulase.

Colourimetric method for the determination of 
endo
-1,4-β-glucanase (cellulase) in enzyme preparations and fermentationproducts

Principle:
(endo-1,4-β-glucanase)
(1) 3-Ketobutylidene-G5-β-PNP + H2O → Blocked-GX + G(5-X)-β-PNP

(thermostable β-glucosidase)
(2) G(5-X)-β-PNP + H2O → D-glucose + PNP

(alkaline solution)
(3) PNP → phenolate ion (yellow colour)
Note: PNP = 4-nitrophenol

Kit size:
K-CELLG5-4V 120 / 240 assays (manual) / 480 (auto-analyser)
or
K-CELLG5-2V 60 / 120 assays (manual) / 240 (auto-analyser)

Method:                         Spectrophotometric at 400 nm
Total assay time:           10 min
Detection limit:                3.5 x 10-4 U/mL
Application examples:
Fermentation broths, industrial enzyme preparations, biofuels research
Method recognition:     Novel method

 

高尔夫球场农药混合标准溶液GF-6


产品编号 产品名称 产品规格 产品等级 产品价格
168-24931 Pesticide Mixture Standard Solution GF-6 (LC) (each 25μg/ml Acetonitrile Solution)
高尔夫球场农药混合标准溶液GF-6
1ml for Pesticide Residue Analysis

高尔夫球场农药混合标准溶液GF-6 (LC) (各25μg/ml丙酮溶液中)

Pesticide Mixture Standard Solution GF-6 (LC) (each 25μg/ml Acetonitrile Solution)

 

成分表(8种成分)


Asulam

磺草灵

Triclopyr

绿草定

Mecoprop (MCPP)

二甲四氯丙酸

Halosulfuron-methyl

氯吡嘧磺隆

Thiuram

秋兰姆

Flazasulfuron

啶嘧磺隆

Siduron

环草隆

Azoxystrobin

嘧菌酯

相关产品


产品编号

产品名

中文名

包装

161-25141

Pesticide   Mixture Standard Solution GF-1 (GC/MS) (each 20μg/ml Acetone Solution)

高尔夫球场农药混合标准溶液GF-1(GC/MS)(各20μg/ml丙酮溶液中)

1ml

166-25211

Pesticide Mixture Standard Solution GF-1 (LC/MS/MS) (each   20μg/ml)

高尔夫球场农药混合标准溶液GF-1 (LC/MS/MS) (各   20μg/ml)

1ml

168-25151

Pesticide Mixture Standard Solution GF-2 (GC/MS) (each 20μg/ml   Acetone Solution)

高尔夫球场农药混合标准溶液GF-2(GC/MS)(各20μg/ml丙酮溶液中)

1ml



去端肽胶原,蜂窝海绵


产品编号 产品名称 产品规格 产品等级 产品价格
KOU-CSH-10 Atelocollagen Honeycomb sponge 100MG
KOU-CSH-96 Atelocollagen Honeycomb Disc 96 25PC


3D培养和3D支架组织工程研究的有用工具去端肽胶原,蜂窝海绵

去端肽胶原,蜂窝海绵

Atelocollagen  Honeycomb

背景

  “蜂窝”胶原海绵具有方向统一、均匀的孔(200-400微米),细胞可以穿透并在其中增殖密集地排列。这种结构有利于营养物质到海绵内为细胞做准备供应,并释放代谢废物和生化产物。细胞能够增殖且填充管腔,形成均匀的细胞团。


去端肽胶原,蜂窝海绵

去端肽胶原蜂窝海绵

Atelocollagen Honeycomb Sponge

(KOU-CSH-10) 为2毫米的立方体,应用于3D细胞

培养物和高密度细胞培养基组织工程细胞支架


去端肽胶原,蜂窝海绵

去端肽胶原蜂窝海绵

Honeycomb Disk 96

(KOU-CSH-96)直径为6毫米圆盘形,

适用于96孔和高通量筛选细胞培养。


去端肽胶原,蜂窝海绵

KOU-CSH-10 : stereoscopic microscope image

立体显微镜图像


去端肽胶原,蜂窝海绵

KOU-CSH-96 : stereoscopic microscope image

立体显微镜图像


去端肽胶原,蜂窝海绵

Electron microscope image of Honeycomb sponge

蜂窝海绵的电子显微图像


去端肽胶原,蜂窝海绵

Electron microscope image of mouse fibroblast

cell culture in 'Honeycomb collagen sponge

蜂窝胶原海绵小鼠成纤维细胞培养的电子显微镜图像

去端肽胶原的特点

  去端肽胶原是由蛋白酶溶解的胶原,但是它的物理性质几乎与天然未加溶的胶原蛋白相同。而去端肽胶原更具有优越的特性。



去端肽胶原,蜂窝海绵

特点与优点

  “蜂窝”胶原海绵由高度纯化I型去端肽胶原制备(牛皮来源),并且可以通过胶原酶降解。

◆应用

  3D培养

  组织工程3D支架研究

◆使用实例

实例 1

NG1RGB人成纤维细胞在蜂窝圆盘(KOU-CSH-96)

去端肽胶原,蜂窝海绵

饲养层细胞/孔(×103

附着细胞于蜂窝圆盘96(×103

10

2.2

20

5.3

40

8.6

细胞增殖:如图所示,在蜂窝圆盘96培养孔中接种。

细胞增殖通过NADH依赖性燃料(WST-8)测定OD值。

细胞附着:圆盘培养如图所示,培养一天后转移至新培养孔,并测定细胞数,显示20%的细胞附着。

实例 2

胚体在蜂窝海绵支架体内移植后的胚体致畸移植

  小鼠肾脏在海绵蜂窝支架移植(EB/+ CSH)胚体移植后12周后,没有形成畸胎瘤的迹象,而无KOU-CSH-10(ES/-CSH)的所有小鼠胚体培养产生畸胎瘤。组织学上,移植ES /+ CSH与相邻的主肾组织没有明显区别,表明没有具体的诱导自发分化难以区分。

  方法:板中培养小鼠胚胎干(ES)细胞进行胰蛋白酶处理,通过尼龙网过滤,接种到96孔板(1×10^4细胞/孔),培养5天,形成胚状体(EB)。当EB与蜂窝海绵(KOU-CSH-10)混合,EB迅速、均匀融入KOU-CSH-10的矩阵中。EB /+ CSH复合物移植到6周龄小鼠的肾筋膜。


去端肽胶原,蜂窝海绵


实例 3

  体内小鼠胚胎干/间质细胞镶嵌球移植KOU-CSH-10支架后新生毛发。
  

  方法:小鼠ES细胞和小鼠胚胎间充质细胞(MDU1)共培养以产生两种细胞的镶嵌球体。镶嵌球体与KOU-CSH-10去端肽胶原蜂窝海绵混合,在6周龄小鼠的背部肌肉移植。

去端肽胶原,蜂窝海绵

<参考文献>
1. Suzuki T, et al. Growth inhibition and differentiation of cultured smooth muscle cells depend on cellular crossbridges across the tubular lumen of type I collagen matrix honeycombs. (2009) Microvasc Res. 77(2):143-149.

相关产品

细胞培养胶原:AteloCell

Atelocollagen, Native collagen

去端肽胶原,天然胶原
Atelocollagen powder

去端肽胶原粉末 

组织培养胶原溶液是KOKEN(东京)生产的高纯度的胶原溶液,先进的实验室生成,优良的品质控制。
[KOU-IPC-30, KOU-IPC-50, IAC-30, IAC-50, KOU-CLP-01]


Atelocollagen, Eagle's MEM, Hanks' Medium, DMEM

Atelocollagen, Eagle's MEM, DMEM 和 RPMI是用于培养的高纯度胶原溶液。
[KOU-MEN-02, KOU-DME-02, KOU-DME-02H, KOU-RPM-02]


Collagen microspheres

胶原微球

胶原微球用来源于牛皮的I型端肽胶原制备的细胞培养基材。该产品可用于培养,如成纤维细胞,上皮细胞和成骨细胞,并已被证明在细胞培养物的长期维持有效。
[KOU-MIC-00]


Atelocollagen,  Honeycomb sponge for cube-shaped and 96-well plate

去端肽胶原,蜂窝海绵立方体和96孔板

蜂窝”胶原海绵由高纯度的牛皮来源I型去端肽胶原制备,并且可以通过胶原酶降解。
[KOU-CSH-10, KOU-CSH-96]


Atelocollagen sponge, Collagen sponge for 35mm culture dish and <90mm × 80mm × 5mm>

去端肽胶原海绵,35mm培养盘与<90mm × 80mm × 5mm >胶原海绵

胶原海绵为3D细胞培养开发的一种胶原的产品。
[KOU-CS-35,KOU-CLS-01]


Atelocollagen sponge, MIGHTY

去端肽胶原海绵,MIGHTY

MIGHTY是强力的胶原海绵,即使施加30kPa(单次)的压缩负荷也不会崩溃。
[KOU-CSM-25, KOU-CSM-50]


Atelocollagen, Permeable membrane for 50mm culture dish

去端肽胶原,50mm培养皿半透膜
Atelocollagen membrane

去端肽胶原膜

胶原膜用高纯度牛皮来源I型去端肽胶原特别制备,用于单层和双层组织培养的研究。
[KOU-MEN-01,KOU-CLF-01]


Atelocollagen, Permeable membrane for 6-well,24-well culture plate, Atelocollagen membrane

去端肽胶原,6孔,24孔培养板,去端肽胶原膜半透膜

由于膜透明,培养时可用显微镜观察细胞。
[KOU-CM-6, KOU-CM-24, KOU-CLF-01]


Type II Collagen II型胶原
Usefull for tissue and cell culture可用于组织和细胞培养[KOU-CL-22]


Atelocollagen coated BETA-TCP scaffold

去端肽胶原涂层BETA-TCP支架
可用于成骨细胞研究[KOU-ACB-05S]

去端肽胶原,蜂窝海绵

AteloCell 细胞培养胶原.pdf

AteloCell® 系列常见问题(FAQ)

AteloCell® 系列非常适合从日常细胞维持到再生医学基础研究的细胞培养



去端肽胶原,蜂窝海绵



◆脂肪变性胶原海绵35mm培养皿


Q1:胶原蛋白海绵是否耐热? 它能承受的最高温度是多少? 当温度升高到高于体温的温度时,它会降解吗?
A1:我们没有测量这种产品的耐热性。以下信息供您参考,由于液体胶原变性在40°C左右,海绵型胶原可能

A1:不会变性,除非达到更高的温度。


Q2:这种胶原蛋白的弹性是什么? 它会容易撕裂吗? 它能承受多少重量? 最重要的是,在处理胶原时,我们

Q2:应该注意哪些?
A2:我们没有测试这种产品的弹性。然而,它可能抵抗一定水平的负载,因为这种产品是冻干的不溶性胶原蛋

A2:白。


Q3:它是否适合移植,例如皮肤移植用于伤口愈合?
A3:这不适合移植,因为是没有消除端肽的天然胶原。


Q4:胶原蛋白海绵会溶解吗? 他们如何溶解? 大概需要多长时间才能使它们完全溶解,特别是移植(如皮肤

Q4:胞)到/入动物模型后? 当胶原溶解时,培养的细胞会发生怎么样的变化?
A4:本产品由不溶性胶原蛋白制成,因此不易溶解。在体内情况下,它会在MMP中溶解。虽然该产品不适合

A4:移植,但有报道称该产品用于体内实验。 根据报告,胶原在8周后从活体中移除。移植胶原被认为将可以

A4:取代移植细胞的胞外基质,直到其消失。


Q5:我们可以用手术刀手动切割胶原片到更小的尺寸吗? 在切割过程中和切割后会造成胶原片/结构/完整性的

Q5:破坏/扭曲/分散吗?
A5:你可以切割胶原板,但如果你使用钝刀,孔结构可能会皱起来。


Q6:细胞会以多大强度/完好地吸附到胶原海绵? 即使在强烈搅拌后细胞能否容易分离?
A6:即使在强烈搅拌后,一旦细胞附着,也不容易分离,除非低粘附性细胞。


Q7:我们如何能够以最小损伤从胶原海绵中分离细胞?
A7:请使用胶原酶分离胶原。

◆胶原海绵,90*80*5mm


Q1:吸水后变形的胶原海绵的厚度是多少?
A1:它将变得略小于5mm。


Q2:一旦吸收水,海绵将变形,除非进行交联。 这方面的交联是什么意思?
A2:考虑通过UV或γ辐射的物理交联或通过交联剂的化学交联。


Q3:胶原海绵是否耐热? 它能承受的最高温度是多少? 当温度升高到高于体温的温度时,它会降解吗?
A3:我们没有测量这种产品的耐热性。以下信息供您参考,由于液体胶原变性在40°C左右,海绵型胶原可能不

A3:会变性,除非达到更高的温度。


Q4:这种胶原蛋白的弹性是什么? 它会容易撕裂吗? 它能承受多少重量? 最重要的是,在处理胶原时,我们

Q4:应该注意哪些?
A4:我们没有测试这种产品的弹性。然而,它可能抵抗一定水平的负载,因为这种产品是冻干的不溶性胶原蛋

A4:白。


Q5:它是否适合移植,例如皮肤移植用于伤口愈合?
A5:这不适合移植,因为是没有消除端肽的天然胶原。


Q6:胶原蛋白海绵会溶解吗? 他们如何溶解? 大概需要多长时间才能使它们完全溶解,特别是移植(如皮肤

Q6:细胞)到/入动物模型后? 当胶原溶解时,培养的细胞会发生怎么样的变化?
A6:本产品由不溶性胶原蛋白制成,因此不易溶解。在体内情况下,它会在MMP中溶解。虽然该产品不适合

A6:移植,但有报道称该产品用于体内实验。 根据报告,胶原在8周后从活体中移除。移植胶原被认为将可以

A6:取代移植细胞的胞外基质,直到其消失。


Q7:我们可以用手术刀手动切割胶原片到更小的尺寸吗? 在切割过程中和切割后会造成胶原片/结构/完整性的

Q7:破坏/扭曲/分散吗?
A7:你可以切割胶原板,但如果你使用钝刀,孔结构可能会皱起来。


Q8:细胞会以多大强度/完好地吸附到胶原海绵? 即使在强烈搅拌后细胞能否容易分离?
A8:即使在强烈搅拌后,一旦细胞附着,也不容易分离,除非低粘附性细胞。


Q9:我们如何能够以最小损伤从胶原海绵中分离细胞?
A9:请使用胶原酶分离胶原。

◆胶原,可渗透膜


Q1:膜是否耐热? 它能承受的最高温度是多少? 当温度升高到高于体温的温度时,它会降解吗?
A1:我们没有测量这种产品的耐热性。以下信息供您参考,由于液体胶原变性在40°C左右,海绵型胶原可能不会

A1:变性,除非达到更高的温度。


Q2:膜的弹性是什么? 它会容易撕裂吗? 它能承受多少重量? 最重要的是,在处理膜时,我们应该注意哪些?
A2:我们没有测试这种产品的弹性。然而,它会容易撕裂,因为这种产品被再加工成薄膜形式。


Q3:它是否适合移植,例如皮肤移植用于伤口愈合?
A3:是的。


Q4:渗透膜会溶解吗? 他们如何溶解? 它需要多长时间才能使它们完全溶解,特别是移植(皮肤细胞)到动物

Q4:模型后? 当胶原溶解时,培养的细胞会发生哪些变化?
A4:我们认为可透膜在移植后约一个月会溶解。


Q5:我们可以用手术刀手动切割膜到更小的尺寸吗? 它会在切割过程中和切割后引起膜结构/完整性的破坏/变

Q5:形/分散吗?
A5:是的,你可以将膜切成更小的尺寸。


Q6:细胞粘附/附着到可渗透膜上的强度/完整性是什么样的,特别是当我们在膜的双面上进行两种不同细胞类型

Q6:的夹心培养时? 即使在强烈搅拌后细胞是否容易分离?
A6:即使在强烈搅拌后,一旦它们附着,细胞也不容易分离,除非低粘附性细胞。


Q7:我们如何能够从双侧膜以最小的损伤分离细胞?
A7:请用刮刀或胶原酶回收细胞。


Q8:如何在膜的两个不同表面上观察两种不同的细胞类型? 我们用镊子翻转? 这种行为是否会造成细胞损伤或

Q8:脱落?
A8:您可以通过相差显微镜观察细胞。然而,难以区分细胞接种于哪一侧。因此,更好的方法是用荧光素标记细

A8:胞并通过荧光显微镜观察。


Q9:可以将膜从50mm培养皿,6孔和24孔培养板上分离下来吗?
A9:可用刀把它分开。

◆胶原,蜂窝海绵


Q1:如何确保培养的细胞粘附在蜂窝海绵上? 我们可以在显微镜下观察吗?
A1:相差显微镜能够观察。


Q2:蜂窝海绵是否耐热? 它能承受的最高温度是多少? 当温度升高到高于体温的温度时,它会降解吗?
A2:我们没有测量这种产品的耐热性。以下信息供您参考,由于液体胶原变性在40°C左右,海绵型胶原可能

A2:不会变性,除非达到更高的温度。


Q3:蜂窝海绵的耐久性是什么? 它能承受多少重量?
A3:我们没有测试这种产品的弹性。然而,它会被负载打破,因为这种产品是冻干低浓度胶原。


Q4:蜂窝海绵会溶解吗? 他们如何溶解? 它需要多长时间才能使它们完全溶解,特别是移植(如皮肤细胞)

Q4:到/入动物模型后? 当蜂窝海绵溶解时,培养细胞会发生哪些变化?
A4:移植后约一个月,海绵会溶解。


Q5:我们可以用手术刀手动切割蜂窝海绵到更小的尺寸(更薄)吗? 在切割过程中和切割后,是否会导致海

Q5:绵结构的破坏/变形/分散?
A5:你可以切割胶原板,但如果你使用钝刀,孔结构可能会皱起来。


Q6:细胞粘附/附着到蜂窝海绵上的强度/完整性如何?即使在强烈搅拌后细胞是否容易分离?
A6:即使在强烈搅拌后,一旦它们附着,细胞也不容易分离,除非低粘附性细胞。


Q7:可以通过胶原酶处理,简便地收获细胞。处理后会影响细胞活力吗?有何处理方案?
A7:请加胶原酶至终浓度为0.1%,溶解约30分钟。如果你担心细胞损伤,提高胶原酶浓度和减少处理时间。


◆羊毛脂海绵(MIGHTY)


Q1:MIGHTY海绵是否耐热? 它能承受的最高温度是多少? 当温度升高,例如高于体温时,它会降解吗?
A1:我们没有测量这种产品的耐热性。以下信息供您参考,由于液体胶原变性在40°C左右,海绵型胶原可能

A1:不会变性,除非达到更高的温度。


Q2:MIGHTY海绵会溶解吗? 他们如何溶解? 它需要多长时间才能使它们完全溶解,特别是移植(如皮肤细

Q2:胞)到/入动物模型后? 当MIGHTY溶解时,培养的细胞会发生哪些变化?
A2:因为MIGHTY海绵是高强度海绵,它难以溶解。有一个数据表明MIGHTY海绵移植后至少3个月内不会溶解。


Q3:我们可以用手术刀手动切割MIGHTY海绵到更小的尺寸吗? 在切割过程中和切割之后,是否会导致MIGHTY

Q3:结构的破裂/变形/分散? 在切割过程中是否有任何的推荐步骤或预防措施?
A3:当它是膨胀状态,你可以切割MIGHTY海绵。然而,刀切割海绵后,可能会有切口,因为这是一种高强度的海绵。


Q4:细胞粘附/附着到MIGHTY海绵上的强度/完整性?即使在强烈搅拌后细胞也能很容易分离吗?
A4:即使在强烈搅拌后,一旦它们附着,细胞也不容易分离,除非低粘附性细胞。


Q5:我们如何将细胞以最小损伤从MIGHTY海绵中分离?
A5:因为MIGHTY海绵是高强度的,所以难以从海绵中回收活细胞。另一方面,在均质化之后回收核苷酸或蛋白质

A5:是可行的。


◆胶原微球


Q1:微球胶原蛋白的上清液/溶液是什么?
A1:PBS


Q2:如何确保培养的细胞粘附到微球? 我们可以在显微镜下观察吗?
A2:你可以通过相差显微镜观察。


◆胶原涂层B-TCP(B-磷酸钙)支架


Q1:这个支架的厚度是多少?
A1:1.0±0.1 mm


Q2:支架是否耐热? 它能承受的最高温度是多少?当温度升高,例如高于体温时,它会降解吗?
A2:β-TCP支架可以耐高温,但是涂覆的胶原在40℃左右变性。


Q3:支架的弹性如何? 它会容易撕裂/破裂吗? 它能承受多少重量? 最重要的是,在处理胶原时,我们应该注

Q3:意哪些?
A3:我们没有测试这种产品的弹性。然而,我们认为支架是来自β-TCP,它可以承受一定负载。


Q4:它适合移植吗?
A4:是。 但本产品设计用于骨形成测定。


Q5:支架会溶解吗? 他们如何溶解? 需要多长时间才能使它们完全溶解,特别是移植到动物模型之后? 当支架

Q5:溶解时,培养的细胞会发生哪些变化?
A5:很难溶解,因为这个产品是由β-TCP组成。


Q6:我们可以用手术刀手动切割支架到更小的尺寸吗? 在切割过程中和切割后会引起支架结构的破坏/变形/分散

Q6:吗?
A6:很难切割,因为这个产品是由β-TCP组成。


Q7:细胞粘附/附着到支架上的强度/完整性如何?即使在强烈搅拌后细胞是否容易分离?
A7:即使在强烈搅拌后,一旦细胞附着,也不容易分离,除非低粘附性细胞。


Q8:我们如何能够以最小的损害从支架上分离细胞?
A8:请使用胶原酶分离胶原。

Atelocollagen, Honeycomb sponge

Product number : KOU-CSH-10, KOU-CSH-96


<References>

1.  Ishii  I,  et  al.  Correlation  between  antizyme  1  and  differentiation  of  vascular  smooth  muscle 

     cellscultured in honeycomb-like type-I collagen matrix. Amino Acids. (2012) Feb;42(2-3):565-75.

2.  Ishii I, et al. Histological and functional analysis  of vascular smooth muscle cells in a novel  culture

     system with honeycomb-like structure. At herosclerosis. (2001) Oct;158(2):377-84.

3.  Mariko Yamaki: in vitro and de novo generation of hair from mosaic spheres formed jointly from ES

     and  mesenchymal  cells.  The  Japanese  Society  for  Regenerative  Medicine magazine.  (2009)

     8(2):91-97.

4.  Mariko  Yamaki:  Artificial  extracellular  matrix  of  type  I  collagen  can  suppress the tumorigenetic

     potential  of  mouse  embryonic  stem  cells.  The  Japanese  Society  for  Regenerative  Medicine

     magazine. (2009) 8(1):109-114.

5.  Suzuki  T,  et  al.  Growth  inhibition  and  differentiation  of  cultured  smooth  muscle  cells  depend 

     on cellular crossbridges across the tubular lumen of type I collagen matrix honeycombs. (2009)

     Microvasc Res. 77(2):143-149.

6.  Fukui N, et al. Bone tissue reaction of nano-hydroxyapatite/collagen composite at the early stage of

     implantation. (2008) Biomed Mater Eng. 18(1):25-33.

7.  Fukushima K, et al. The axonal regeneration across a honeycomb collagen sponge applied to the

     transected  spinal cord. (2008) J Med Dent Sci. 55(1):71-79.

8.  Kakudo N, et al. Bone tissue engineering using human adipose-derived stem cells and honey  comb

     collagen scaffold. (2008) J Biomed Mater Res A. 84(1):191-197.

9.  Saeki K, et al. Highly efficient and feeder-free production of subculturable vascular endothelial  cells

     from primate embryonic stem cells.(2008) J Cell Physiol. 217(1):261-280

10.Takeuchi R, et al. Low-intensity pulsed ultrasound activates the phosphatidylinositol 3 kinase/Akt

     pathway and stimulates the growth of chondrocytes in three-dimensional cultures: a basic science

     study. (2008) Arthritis Res Ther. 10(4):R77.

11.Hidetsugu T, et al. Mechanism of bone inducti on by KUSA/A1 cells using atelocollagen honeycomb

     scaffold. (2007) J Biomed Sci. 14(2):255-263.

12.Rodriguez  AP,  Missana  L,  Nagatsuka  H,  et  al.:  Efficacy  of  atelocollagen  honeycomb  scaffold 

     in bone formation using KUSA/A1 cells. J Biomed Mater Res A. (2006) 77(4):707-717.

13.George J, et al. Differentiation of mesenchymal stem cells into osteoblasts on honeycomb collagen

     scaffolds. (2006) Biotechnol Bioeng. 95(3):404-411.

14.Imamura  T,  et  al.  Embryonic  stem  cell-derived  embryoid  bodies  in  three-dimensional  culture

     system form hepatocyte-like cells in vitro and in vivo. (2004) Tissue Eng. 10(11-12):1716-1724.

15.Itoh H, et al. A honeycomb collagen carrier for cell culture as a tissue engineering scaffold.(2001) Artif

     Organs. 25(3):213-217.

16.Moriyama  T,  et  al.  Development  of  composite  cultured  oral  mucosa  utilizing  collagen  sponge

     matrix and contracted collagen gel: a preliminary study for clinical applications. (2001) Tissue Eng.7(4):

     415-427.

L-苹果酸[液体即用型]检测试剂盒 L-Malic Acid Assay Kit (Liquid Ready Reagents) 货号:K-LMALQR Megazyme中文站

L-苹果酸[液体即用型]检测试剂盒

英文名:L-Malic Acid Assay Kit (Liquid Ready Reagents)

货号:K-LMALQR

规格:1100 assays (microplate) / 1100 (auto-analyser)

The L-Malic Acid (Liquid Ready Reagents) test kit is a rapid, simple, reliable and accurate method for the specific measurement and analysis of L-malic acid in wine, beverages, foodstuffs and other materials. Supplied as a “ready to use” liquid stable formulation that is suitable for auto-analyser and microplate formats.
Suitable for auto-analyser and microplate formats.

UV-method suitable for microplate and auto-analyser formats
for the determination of L-Malic Acid in foodstuffs, beverages
and other materials

Principle:
(L-malate dehydrogenase)
(1) L-Malic acid + NAD+ ↔ oxaloacetate + NADH + H+

(glutamate-oxaloacetate transaminase)
(2) Oxaloacetate + L-glutamate → L-aspartate + 2-oxoglutarate

Kit size: 1100 assays (microplate)
/ 1100 (auto-analyser)
Method: Spectrophotometric at 340 nm
Reaction time: ~ 3 min
Detection limit: 166 mg/L (recommended format)
Application examples:
Wine, beer, fruit juices, soft drinks, candies, fruit and vegetables,
bread, cosmetics, pharmaceuticals and other materials (e.g. biological
cultures, samples, etc.)
Method recognition:
Methods based on this principle have been accepted by AOAC, EEC,
EN, NF, NEN, DIN, GOST, OIV, IFU, AIJN and MEBAK

Advantages

  • PVP incorporated to prevent tannin inhibition
  • “Ready to use” liquid stable formulation
  • Very competitive price (cost per test)
  • All reagents stable for > 18 Months
  • Very rapid reaction (~ 3 min)
  • Standard included
  • Suitable for microplate and auto-analyser formats

Q1. Should the pH of the sample be adjusted even for samples in acidic media?

The pH of the assay solution after the sample is added should be the same as that of the assay buffer that is supplied with the kit.
Low sample volumes (e.g. 0.1 mL) are not likely to affect the pH of the assay solution and therefore may not require pH adjustment.
Samples above 0.1 mL are more likely to affect the pH of the assay solution and therefore the pH of these samples should be adjusted as described in the data booklet, prior to addition to the assay.

Q2. Sometimes a negative absorbance change is obtained for the blank samples, is this normal? Should the real value (negative absorbance change) or “0” be used in the calculation of results?

Sometimes the addition of the last assay component can cause a small negative absorbance change in the blank samples due to a dilution effect and in such cases it is recommended that the real absorbance values be used in the calculation of results.

Q3. There is an issue with the performance of the kit; the results are not as expected.

If you suspect that the Megazyme test kit is not performing as expected such that expected results are not obtained please do the following:

  1. Ensure that you have tested the standard sample that is supplied with the Megazyme test kit.
  2. Send the results of the kit standard, blank samples and the results obtained for your sample, in the relevant MegaCalc spreadsheet (if available) to Megazyme (cs@megazyme.com). Where available the relevant MegaCalc spreadsheet can be downloaded from where the product appears on the Megazyme website.
  3. State the kit lot number being used (this is found on the outside of the kit box).
  4. State which assay format was used (refer to the relevant page in the kit booklet if necessary).
  5. State exact details of any modifications to the standard procedure that is provided by Megazyme.
  6. State the sample type and describe the sample preparation steps if applicable.

Q4. What is the difference between K-LMAL-58A / 116A, K-LMALAF, K-LMALMQ and K-LMALQR?

Megazyme produces 4 L-malic acid test kits:
K-LMAL-58A / 116A: UV method, automated format for use with auto-analysers.
K-LMALAF: UV method, manual format for use with spectrophotometers.
K-LMALMQ: Colourimetric method, manual format for use with hand held colorimeter.
K-LMALQR: UV method, liquid ready reagents automated format for use with auto-analysers.

Q5. Which L-Malic Acid Kit is recommended for a 96-well microplate format?

Auto-analysers use ~ 0.315 mL reaction volumes and pathlengths between 4-8 mm which is similar to a standard 96-well microplate where a 0.315 mL reaction volume would give a pathlength of ~ 6-7 mm.  Therefore, K-LMALAF can be used directly in a 96-well microplate format with minimal assay optimisation.
If preferred, K-LMAL-58A / 116A may also be easily converted for use in a 96-well microplate format.  Basically, the assay volumes for the cuvette format must be reduced approximately 10-fold for use in a 96-well microplate. However, some assay optimisation may be required (e.g. increased enzyme concentration etc.) and unlike the cuvette which has a set pathlength of 1 cm, the pathlength in the microplate is dependent upon the volume of liquid in the well.
Therefore to enable the calculation of the amount of analyte in the samples from tests performed in the microplate format one of the following must be done:

  1. The easiest method is to use a microplate reader that has a pathlength conversion capability (i.e. the microplate reader can detect the pathlength of each well and convert the individual readings to a 1 cm pathlength).  This will allow values to be calculated using the MegaCalc calculation software which can be found where the product is located on the Megazyme website.
  2. Perform a standard curve of the analyte on each microplate that contains test samples and calculate the result of the test samples from the calibration curve (concentration of analyte versus absorbance).
  3. Perform a standard curve of the analyte in both the cuvette format (i.e. with a 1 cm pathlength) and the 96-well microplate format and use these results to obtain a mean conversion factor between the cuvette values and the microplate values.

L-Malic Acid Kit Recommendation For Microplate Format:
Either K-LMAL-58A / 116A or K-LMALAF is recommended for use in a 96-well microplate format and the main advantages / disadvantages are described below:
K-LMAL-58A / 116A:
The assay volumes of this kit should be reduced by 10-fold for use in a 96-well microplate format (some assay optimisation may be required, e.g. increased enzyme concentration etc.).
The calculation of results is achieved as outlined above in either of points 1, 2 or 3.
The main advantage here is that if this kit is used with a microplate reader that has a pathlength conversion capability, or if results are converted as outlined above in point 3, then this enables easy calculation of results using the K-LMAL-58A / 116A MegaCalc application (available on the Megazyme website where the product is located).
K-LMALAF:
This kit is designed for use in an auto-analyser and therefore can be used without any modification to assay volumes directly in a 96-well microplate format. 
This kit has less reagent additions than K-LMAL-58A / 116A.
K-LMALAF does not have a MegaCalc application available to enable easy results calculation which therefore must be achieved as outlined above in either of points 2 or 3.

Q6. Do samples require any specific sample preparation prior to testing with the kits?

The sample preparation is sample dependent, some samples may be tested directly in the assay or after appropriate dilution, however, some samples may require further sample preparation prior to testing.  The following are example of sample preparation methods:
(a) Liquid samples: clear, slightly coloured and approximately neutral, liquid samples can be used directly in the assay.
(b) Acidic samples: if > 0.1 mL of an acidic sample is to be used undiluted (such as wine or fruit juice), the pH of the solution should be increased to approx. 9.0 using 2 M NaOH, and the solution incubated at room temperature for 30 min.
(c) Carbon dioxide: samples containing significant quantities of carbon dioxide, such as beer, should be degassed by increasing the pH to approx. 9.0 with 2 M NaOH and gentle stirring, or by stirring with a glass rod.
(d) Coloured samples: an additional sample blank, i.e. sample with no L-MDH, may be necessary in the case of coloured samples.
(e) Strongly coloured samples: if used undiluted, strongly coloured samples should be treated by the addition of 0.2 g of PVPP/10 mL of sample.  Shake the tube vigorously for 5 min and then filter through Whatman No. 1 filter paper.
(f) Solid samples: homogenise or crush solid samples in distilled water and filter if necessary.
(g) Samples containing fat: extract such samples with hot water at a temperature above the melting point of the fat, e.g. in a 100 mL volumetric flask.  Adjust to room temperature and fill the volumetric flask to the mark with distilled water.  Store on ice or in a refrigerator for 15-30 min and then filter.  Discard the first few mL of filtrate, and use the clear supernatant (which may be slightly opalescent) for assay. Alternatively, clarify with Carrez reagents.
(h) Samples containing protein: deproteinise samples containing protein by adding an equal volume of ice-cold 1 M perchloric acid with mixing.  Centrifuge at 1,500 g for 10 min and neutralise the supernatant with 1 M KOH.  Alternatively use Carrez reagents.

Q7. How can I work out how much sample to extract and what dilution of my sample should be used in the kit assay?

Where the amount of analyte in a liquid sample is unknown, it is recommended that a range of sample dilutions are prepared with the aim of obtaining an absorbance change in the assay that is within the linear range.
Where solid samples are analysed, the weight of sample per volume of water used for sample extraction/preparation can be altered to suit, as can the dilution of the extracted sample prior to the addition of the assay, as per liquid samples.

Q8. Can you explain, step by step, how to follow the method and perform the kit assay?

For users who are not familiar with how to use the Megazyme tests kits then it is recommended that they follow this example, e.g. D-Fructose/D-Glucose Assay kit K-FRUGL (http://secure.megazyme.com/D-Fructose-D-Glucose-Assay-Kit):

1. The kit components are listed on pages 2-3 of the kit booklet.
2. Prepare the kit reagents as described on page 3.
3. For separate measurements of glucose and fructose follow procedure A on page 4.
4. Pipette the volumes listed for water, sample, solution 1 and solution 2 into 3 mL, 1 cm pathlength cuvettes. Duplicate sample assays and duplicate blanks are recommended. Mix the contents of each cuvette by inversion (seal the cuvette using parafilm or a plastic cuvette cap – do not use a finger) then after ~3 min record the first absorbance reading of each cuvette at 340 nm (this is reading A1).
5. Then add suspension 3 and mix the contents of each cuvette by inversion. Incubate for 5 minutes then record the absorbance reading of each cuvette at 340 nm (this is reading A2). NB. It is essential that the reaction is compete. To assess this, record the absorbances at ~ 2 minute intervals and until the absorbance plateaus. A stable absorbance indicates that the reaction is complete. If the absorbance continues to increase then continue to record absorbances until it plateaus and only then record absorbance reading A2.
6. Then add suspension 4 and mix the contents of each cuvette by inversion. Incubate for 5 minutes then take absorbance reading of each cuvette at 340 nm (this is reading A3). NB. As above, assess that the reaction has completed by take subsequent readings at ~2 min intervals.
7. For simple, automated results analysis, input the absorbance readings (A1, A2, A3) for samples and blanks into the 
K-FRUGL MegaCalc.

To ensure that the assay is working, and being performed correctly it is recommend that the test is performed using the standard sample that is provided with the kit and to obtain the expected values before proceeding to test real samples.
It is recommend that new users also watch 
this video which highlights how to perform the assays.
Many of the other Megazyme test kits follow a similar format.

Q9. Can the sensitivity of the kit assay be increased?

视频

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