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首页 > 蛋白生物学 > 蛋白Marker > 多肽电泳Marker

超低分子量蛋白Marker III(3.4-100kd)(非预染)

超低分子量蛋白Marker III(3.4-100kd)(非预染)

产品编号:RTD6125

产品规格:10次(50μl)

数量
价格 ¥300

超低分子量蛋白质Marker III3.4-100kD

Ultra Low Molecular Weight Protein Marker III

产品编号及规格:

RTD6125   10T50μl)   300元

储存条件:

  -20保存,开封后有效期12个月。

产品简介:

本产品包含5种多肽和3种低分子量蛋白质组成,分子量范围为3.4kD-100kD。可以用来判断 SDS-PAGE上多肽和小蛋白的分子量。  


使用说明:

第一次收到该产品,室温融化后,彻底混匀,离心快甩将溶液完全收集到管底,根据需要适量分装成小管,-20℃贮存,每次取一小管使用;本产品为即用型,融化后既能使用,不能95加热处理

. 制胶:

I 配制分离胶

1.按照表一将不同体积的双蒸水、40%PAA19:1)、凝胶缓冲液和乙二醇加入到小烧杯中混合。

2.加入10%APSTEMED,立即混匀5-10秒,以使溶液充分混匀。

3.在凝胶模具中迅速灌入适量分离胶溶液,然后在分离胶溶液上轻轻覆盖一层1-3 cm的水层,使凝胶表面保持平整。

4.静置30-60分钟,待分离胶和水层之间出现一个清晰的界面表示凝胶已聚合。

表一  (一块0.75mm mini胶用量)

 

分离胶

浓缩胶

 

18T, 5%C /6.0 ml

5T, 3.3%C/2 ml

40% PAA19:1

2.7 ml

/

40% PAA (29:1)

/

0.25 ml

4×凝胶缓冲液

1.5 ml

0.5 ml

乙二醇(电泳级)

1.8 ml

/

ddH2O

/

1.25 ml

10APS

50-65 μl

20 μl

TEMED

6 μl

2 μl

注:如非必须,不要使用1.0mm和1.5mm的凝胶,尽量使用厚度0.75mm的凝胶,这样会减少电泳后染色和脱色的时间。

II 配制浓缩胶

去除覆盖在分离胶上的水层,用滤纸将残留的水吸去。

1.按照表一将不同体积的双蒸水、40%PAA(29:1)和凝胶缓冲液加入到小烧杯中混合。

2.加入10%过硫酸铵和TEMED,立即混匀5-10秒,以使溶液充分混匀。

3.将梳子插入凝胶内,避免产生气泡。

4.静置30-60分钟装待凝胶聚合

. 电泳:

1. 取一管分装好的Marker样品,彻底融化,不要加热处理,充分混匀后备用。

2. 电泳前,将10×Tris-Tricine-SDS电泳缓冲液(Cat No:CB010)用蒸馏水稀释成1×缓冲液备用。将电泳槽的内外槽加入1×缓冲液,轻柔拔出梳子,将Marker或蛋白样品(样品已经过2×Tricine上样缓冲液[Cat No:TP050]处理)加入点样孔,根据表二条件进行电泳,至蓝色指示前沿至分离胶3/4位置时即可停止电泳。整个电泳过程大约需要2-3个小时。注:如果电泳时间过短,染色时,指示前沿容易遮挡小于5kd的小肽。

表二 多肽电泳条件

恒电压

150 V

起始电流

45-55 mA

结束电流

 10-15 mA

电泳时间

2-3小时

. 染色

根据常规实验步骤,用配方5和6(表三)进行染色和观察。如果使用配方5进行染色时效果不好或考虑其毒性,请选择本公司的FastBlue蛋白染色液(Cat No:RTD6202),该产品能在5分钟之内完成蛋白的染色,不至于小肽在染色和脱色中从凝胶中脱离,是常规染色液的理想替代品。

表三 小分子蛋白质SDS-PAGE电泳试剂配制

1. 40%PAA29:1)(配制浓缩胶)    

   丙稀酰胺       38.67g

   甲叉双丙稀酰胺 1.33g

 ddH2O溶解后定容至100 ml,过滤后使用。

贮存:4

2.  40% PAA19:1 (配制分离胶)

丙稀酰胺      38 g

甲叉双丙稀酰胺 2 g

ddH2O溶解后定容至100 ml,过滤后使用。

贮存:4

3. 4×凝胶缓冲液(配制凝胶用)

[3MTris; 0.4% SDS; pH8.45]

Tris碱 36.3 g;ddH2O 80ml

0.4 g SDS或4 ml 10% SDS

用HCl调pH值至8.45;用ddH2O定容至100 ml

贮存:4℃


4. 10×Tris-Tricine-SDS电泳缓冲液

[1MTris;1MTricine ;1% SDS; pH 8.3]

Tris 121.14g

Tricine 179.2g

SDS10g

ddH2O溶解,定容至1000ml(不用调pH)

贮存:4

注:使用前稀释成缓冲液使用

5. 染色液

冰醋酸            100 ml

甲醇              450 ml

考马斯亮蓝G-250  0.25 g

                450 ml

 

6. 脱色液

冰醋酸  100ml

      900ml

7.  2×Tricine蛋白样品上样缓冲液(10ml

1ml 1MTris-Cl pH6.8

2.4ml 甘油

0.8g  SDS

0.31g DTT(或者400μl β-巯基乙醇)

2mg 考马斯亮蓝G-250

用灭菌ddH2O定容至10ml

混匀分装-20贮存备用

 



多肽电泳配套试剂

名称

货号

规格

超低分子量蛋白电泳试剂盒

RTD6120

10

超低分子量蛋白Marker I3.3-20.1 kD

RTD6101

10次(100 μl

超低分子量蛋白Marker III3.4-100 kD

RTD6125

10次(50 μl

彩虹预染超低分子量蛋白Marker1.2-45 kD

RTD6110

10 50 μl

  彩虹预染超低分子量蛋白Marker II3-40 kD

RTD6145-01

10 50 μl

40% PAA19:1

AC1914-01

100ml

40% PAA29:1

AC2914-01

100ml

多肽电泳凝胶缓冲液

GB010-100ml

100ml

2×Tricine 上样缓冲液(变性,还原)

TP050

10×1ml

FastBlue蛋白染色液

RTD6202

500ml

乙二醇(电泳级)

EA0582-02

100 ml

10×Tris-Tricine-SDS缓冲液(变性电泳,粉末型)

CB010P

500 ml

10×阳极缓冲液 (多肽电泳用

AB080

250 ml

10×阴极缓冲液(多肽电泳用)

CB010

100 ml

 



使用蛋白Marker产品发表部分文章列表:

1. [2021 IF=5.5] Garviecin LG34, a novel bacteriocin produced by Lactococcus garvieaeisolated from traditional Chinese fermented cucumber.

Author: Yurong Gao, Dapeng Li, Shan Liu, Liyuan Zhang

Marker: RTD6110,彩虹超低分子量蛋白Marker(1.2-45 kD)

Journal: Food Control 50 (2015) 896e900

Institution:Key Laboratory of Veterinary Medicine of Heilongjiang Bayi Agricultural University

Paper linkhttp://dx.doi.org/10.1016/j.foodcont.2014.10.040

 

2. [2021 IF=5.5] Purification and partial characterization of M1-UVs300, a novel bacteriocin produced by   isolated from fermented sausage Lactobacillus plantarum.

Author: Yu An, Ying Wang, Xiaoyue Liang, Huaxi Yi, Zhaohang Zuo, Xiaoxi Xu, Dongjie

Zhang, Changqing Yu, Xue Han

Marker: RTD6110,彩虹超低分子量蛋白Marker(1.2-45 kD)

Journal: Food Control  Volume 81, November 2017, Pages 211-217

Institution:Key Laboratory of Veterinary Medicine of Heilongjiang Bayi Agricultural University

Paper linkhttps://doi.org/10.1016/j.foodcont.2017.05.030

 

3. [2021 IF=5.75] The Verticillium dahliae SnodProt1-Like Protein VdCP1 Contributes to Virulence and Triggers the Plant Immune System.

Author: Yi Zhang, Yuhan Gao, Yingbo Liang, Yijie Dong, Xiufen Yang, Jingjing Yuan and Dewen Qiu

Marker: RTD6110,彩虹超低分子量蛋白Marker(1.2-45 kD)

Journal: Frontiers in Plant Science.  Volume 81, November 2017, Pages 211-217

InstitutionInstitute of Plant Protection, Chinese Academy of Agricultural Sciences

Paper linkhttps://doi.org/10.1093/jxb/ery351

 

4. [2021 IF=2.99] Purifcation, characterization, and action mechanism of plantaricin DL3, a novel bacteriocin against Pseudomonas aeruginosa produced by Lactobacillus plantarum DL3 from Chinese Suan Tsai.

Author: Xinran Lv,  Yang Lin ,Yu Jie, Mengtong Sun, Bolin Zhang,Fengling Bai,Hongfei Zhao,Jianrong Li

Marker: RTD6127,超低分子量蛋白Marker IV(3.3-45 kD)

Journal: European Food Research and Technology. Vol 244, pages 323–331 (2018)

InstitutionCollege of Biological Science and Biotechnology, Beijing Forestry University

Paper linkhttps://link.springer.com/article/10.1007/s00217-017-2958-3

 

5. [2021 IF=6.99] Verticillium dahliae Alt a 1-like protein PevD1 targets cotton PR5-like protein and contributes to fungal infection.

Author: Yi Zhang, Yuhan Gao, Yingbo Liang, Yijie Dong, Xiufen Yang, Dewen Qiu

Marker: RTD6110,彩虹超低分子量蛋白Marker (1.2-45 kD)

Journal: Journal of Experimental Botany, Vol 70, Issue 2, 15 January 2019,613-626

InstitutionInstitute of Plant Protection, Chinese Academy of Agricultural Sciences

Paper linkhttps://doi.org/10.1093/jxb/ery351

 

6. [2021 IF=11.277] An  Alternative  Splicing  Variant  of  PtRD26  Delays  Leaf  Senescence by Regulating Multiple NAC Transcription Factors in Populus.

Author: Hou-Ling Wang,Yi Zhang,Ting Wang, Qi Yang,Yanli Yang,Ze Li, Bosheng Li, Xing Wen, Wenyang Li, Weilun Yin, Xinli Xia, Hongwei Guo, and Zhonghai Li

Marker: RTD6110,彩虹预染超低分子量蛋白(1.2-45 kD)

Journal: Plant Cell. Volume 33, Issue 5, May 2021, Pages 1594–1614

InstitutionBeijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing Forestry University

Paper linkhttps://doi.org/10.1093/plcell/koab046

 

7. [2021 IF=2.89] Rapid and sensitive detection ofStaphylococcus aureus and Klebsiella pneumonia based on bacitracin-modified Fe3O4@PDA magnetic beads combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry.

Author: Feng Qin, Shujuan Wang, Mingxia Gao  and  Xiangmin Zhang

Marker: RTD6110,彩虹预染超低分子量蛋白(1.2-45 kD)

Journal: Analytical Methods. 2021,13, 2804-2811

InstitutionDepartment of Chemistry, Fudan University

Paper linkhttps://pubs.rsc.org/en/content/articlelanding/2021/ay/d1ay00614b

 

8. [2021 IF=3.74] Cloning, expression, and function of ferritins in the tick Haemaphysalis flava.

Author: Yu Zhao, Lei Liu, Jin-Bao Liu, Cong-Ying Wu, De-Yong Duan, Tian-Yin Cheng

Marker: RTD6142 高分子量非变性电泳蛋白质Marker II(45-669 kD)

Journal: Ticks and Tick-borne Diseases. Volume 13, Issue 2, March 2022, 101892

InstitutionCollege of Veterinary Medicine, Hunan Agricultural University,

Paper linkhttps://linkinghub.elsevier.com/retrieve/pii/S1877959X21002454

 

9. [2021 IF=0.886] Purification and Characterization of an Intracellular β-Glucosidase from the Protoplast Fusant of Aspergillus oryzae and Aspergillus niger

Author: F.M. Zhu, B. Du, H.S. Gao, C.J. Liu, and J. Lia

Marker: RTD6109 高分子量蛋白质Marker(43-200 kD)

Journal: Applied Biochemistry and Microbiology, 2010, Vol. 46, No. 6, pp. 626–632

InstitutionCollege of Food Science and Technology, Hebei Normal University of Science and Technology

Paper linkhttps://link.springer.com/article/10.1134/S0003683810060116

 

10. [2021 IF=4.374] Effect of Fuzhuan brick-tea addition on the quality and antioxidant activity of skimmed set-type yoghurt.

Author: Dong Liu

Marker: RTD6103 低分子量蛋白质Marker II(14.4-116 kD)

Journal: International Journal of Dairy Technology, Vol 71 March 2018 Pages 22-33

InstitutionDepartment of Food Processing Technology, Anqing Vocational & Technical College

Paper linkhttps://doi.org/10.1111/1471-0307.12395

 

11. [2021 IF=3.7] Smart delivery of poly-peptide composite for effective cancer therapy

Author: Xin Yuan, Yingzhou Qin, Qingmei Tian, Cuijuan Liu, Xiangzhou Meng, Bo Qie, Fan Gao, Ying Huang, Guanghui Xu, Guang Yang, Yimin Zhu

Marker: Protein Marker

Journal: Biomedical Materials, 2022 Jan 24;17(2)

Institution:School of Nano-Tech and Nano-Bionics, University of Science and Technology of China

Paper linkhttps://pubmed.ncbi.nlm.nih.gov/34996052/

 


 
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