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首頁(yè) > 蛋白生物學(xué) > 蛋白Marker > 多肽電泳Marker

超低分子量蛋白Marker III(3.4-100kd)

超低分子量蛋白Marker III(3.4-100kd)

產(chǎn)品編號:RTD6125

產(chǎn)品規格:10次

數量
價(jià)格 ¥300

超低分子量蛋白質(zhì)Marker III3.4-100kD

Ultra Low Molecular Weight Protein Marker III

產(chǎn)品編號及規格:

RTD6125   10T50μl)   300元

儲存條件:

  -20保存,開(kāi)封后有效期12個(gè)月。

產(chǎn)品簡(jiǎn)介:

本產(chǎn)品包含5種多肽和3種低分子量蛋白質(zhì)組成,分子量范圍為3.4kD-100kD??梢杂脕?lái)判斷 SDS-PAGE上多肽和小蛋白的分子量。  


使用說(shuō)明:

第一次收到該產(chǎn)品,室溫融化后,徹底混勻,離心快甩將溶液完全收集到管底,根據需要適量分裝成小管,-20℃貯存,每次取一小管使用;本產(chǎn)品為即用型,融化后既能使用,不能95加熱處理。

. 制膠:

I 配制分離膠

1.按照表一將不同體積的雙蒸水、40%PAA19:1)、凝膠緩沖液和乙二醇加入到小燒杯中混合。

2.加入10%APSTEMED,立即混勻5-10秒,以使溶液充分混勻。

3.在凝膠模具中迅速灌入適量分離膠溶液,然后在分離膠溶液上輕輕覆蓋一層1-3 cm的水層,使凝膠表面保持平整。

4.靜置30-60分鐘,待分離膠和水層之間出現一個(gè)清晰的界面表示凝膠已聚合。

表一  (一塊0.75mm mini膠用量)

 

分離膠

濃縮膠

 

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×凝膠緩沖液

1.5 ml

0.5 ml

乙二醇(電泳級)

1.8 ml

/

ddH2O

/

1.25 ml

10APS

50-65 μl

20 μl

TEMED

6 μl

2 μl

注:如非必須,不要使用1.0mm和1.5mm的凝膠,盡量使用厚度0.75mm的凝膠,這樣會(huì )減少電泳后染色和脫色的時(shí)間。

II 配制濃縮膠

去除覆蓋在分離膠上的水層,用濾紙將殘留的水吸去。

1.按照表一將不同體積的雙蒸水、40%PAA(29:1)和凝膠緩沖液加入到小燒杯中混合。

2.加入10%過(guò)硫酸銨和TEMED,立即混勻5-10秒,以使溶液充分混勻。

3.將梳子插入凝膠內,避免產(chǎn)生氣泡。

4.靜置30-60分鐘裝待凝膠聚合

. 電泳:

1. 取一管分裝好的Marker樣品,徹底融化,不要加熱處理,充分混勻后備用。

2. 電泳前,將10×Tris-Tricine-SDS電泳緩沖液(Cat No:CB010)用蒸餾水稀釋成1×緩沖液備用。將電泳槽的內外槽加入1×緩沖液,輕柔拔出梳子,將Marker或蛋白樣品(樣品已經(jīng)過(guò)2×Tricine上樣緩沖液[Cat No:TP050]處理)加入點(diǎn)樣孔,根據表二條件進(jìn)行電泳,至藍色指示前沿至分離膠3/4位置時(shí)即可停止電泳。整個(gè)電泳過(guò)程大約需要2-3個(gè)小時(shí)。注:如果電泳時(shí)間過(guò)短,染色時(shí),指示前沿容易遮擋小于5kd的小肽。

表二 多肽電泳條件

恒電壓

150 V

起始電流

45-55 mA

結束電流

 10-15 mA

電泳時(shí)間

2-3小時(shí)

. 染色

根據常規實(shí)驗步驟,用配方5和6(表三)進(jìn)行染色和觀(guān)察。如果使用配方5進(jìn)行染色時(shí)效果不好或考慮其毒性,請選擇本公司的FastBlue蛋白染色液(Cat No:RTD6202),該產(chǎn)品能在5分鐘之內完成蛋白的染色,不至于小肽在染色和脫色中從凝膠中脫離,是常規染色液的理想替代品。

表三 小分子蛋白質(zhì)SDS-PAGE電泳試劑配制

1. 40%PAA29:1)(配制濃縮膠)    

   丙稀酰胺       38.67g

   甲叉雙丙稀酰胺 1.33g

 ddH2O溶解后定容至100 ml,過(guò)濾后使用。

貯存:4

2.  40% PAA19:1 (配制分離膠)

丙稀酰胺      38 g

甲叉雙丙稀酰胺 2 g

ddH2O溶解后定容至100 ml,過(guò)濾后使用。

貯存:4

3. 4×凝膠緩沖液(配制凝膠用)

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

Tris堿 36.3 g;ddH2O 80ml

0.4 g SDS或4 ml 10% SDS

用HCl調pH值至8.45;用ddH2O定容至100 ml

貯存:4℃

4. 10×Tris-Tricine-SDS電泳緩沖液

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

Tris 121.14g

Tricine 179.2g

SDS10g

ddH2O溶解,定容至1000ml(不用調pH)。

貯存:4

注:使用前稀釋成緩沖液使用

5. 染色液

冰醋酸            100 ml

甲醇              450 ml

考馬斯亮藍G-250  0.25 g

                450 ml

 

6. 脫色液

冰醋酸  100ml

      900ml

7.  2×Tricine蛋白樣品上樣緩沖液(10ml

1ml 1MTris-Cl pH6.8

2.4ml 甘油

0.8g  SDS

0.31g DTT(或者400μl β-巰基乙醇)

2mg 考馬斯亮藍G-250

用滅菌ddH2O定容至10ml

混勻分裝-20貯存備用

 



多肽電泳配套試劑

名稱(chēng)

貨號

規格

超低分子量蛋白電泳試劑盒

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

彩虹預染超低分子量蛋白Marker1.2-45 kD

RTD6110

10 50 μl

  彩虹預染超低分子量蛋白Marker II3-40 kD

RTD6145-01

10 50 μl

40% PAA19:1

AC1914-01

100ml

40% PAA29:1

AC2914-01

100ml

多肽電泳凝膠緩沖液

GB010-100ml

100ml

2×Tricine 上樣緩沖液(變性,還原)

TP050

10×1ml

FastBlue蛋白染色液

RTD6202

500ml

乙二醇(電泳級)

EA0582-02

100 ml

10×Tris-Tricine-SDS緩沖液(變性電泳,粉末型)

CB010P

500 ml

10×陽(yáng)極緩沖液 (多肽電泳用

AB080

250 ml

10×陰極緩沖液(多肽電泳用)

CB010

100 ml

 



使用蛋白Marker產(chǎn)品發(fā)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,彩虹預染超低分子量蛋白(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,彩虹預染超低分子量蛋白(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 高分子量非變性電泳蛋白質(zhì)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 高分子量蛋白質(zhì)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 低分子量蛋白質(zhì)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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