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超低分子量蛋白質(zhì)Marker III(3.4-100kD)
Ultra Low Molecular Weight Protein Marker III
●產(chǎn)品編號(hào)及規(guī)格:
RTD6125 10T(50μl) 300元
● 儲(chǔ)存條件:
-20℃保存,開封后有效期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??梢杂脕砼袛?SDS-PAGE上多肽和小蛋白的分子量。
● 使用說明:
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(jù)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。
一. 制膠:
I 配制分離膠
1.按照表一將不同體積的雙蒸水、40%PAA(19:1)、凝膠緩沖液和乙二醇加入到小燒杯中混合。
2.加入10%APS和TEMED,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(xiàn)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 |
|
18%T, 5%C /6.0 ml |
5%T, 3.3%C/2 ml |
40% PAA(19:1) |
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(jí)) |
1.8 ml |
/ |
ddH2O |
/ |
1.25 ml |
10%APS |
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(nèi),避免產(chǎn)生氣泡。
4.靜置30-60分鐘裝待凝膠聚合
二. 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(nèi)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2×Tricine上樣緩沖液[Cat No:TP050]處理)加入點(diǎn)樣孔,根據(jù)表二條件進(jìn)行電泳,至藍(lán)色指示前沿至分離膠3/4位置時(shí)即可停止電泳。整個(gè)電泳過程大約需要2-3個(gè)小時(shí)。注:如果電泳時(shí)間過短,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(jié)束電流 |
10-15 mA |
電泳時(shí)間 |
2-3小時(shí) |
三. 染色
根據(jù)常規(guī)實(shí)驗(yàn)步驟,用配方5和6(表三)進(jìn)行染色和觀察。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(qǐng)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(nèi)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(guī)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%PAA(29:1)(配制濃縮膠) 丙稀酰胺 38.67g 甲叉雙丙稀酰胺 1.33g 用ddH2O溶解后定容至100 ml,過濾后使用。 貯存:4℃ |
2. 40% PAA(19:1) (配制分離膠) 丙稀酰胺 38 g 甲叉雙丙稀酰胺 2 g 用ddH2O溶解后定容至100 ml,過濾后使用。 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(diào)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堿 121.14g Tricine 179.2g SDS10g 用ddH2O溶解,定容至1000ml(不用調(diào)pH)。 貯存:4℃ 注:使用前稀釋成1×緩沖液使用 |
5. 染色液 冰醋酸 100 ml 甲醇 450 ml 考馬斯亮藍(lán)G-250 0.25 g 水 450 ml
6. 脫色液 冰醋酸 100ml 水 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(lán)G-250 用滅菌ddH2O定容至10ml 混勻分裝-20℃貯存?zhèn)溆?/span> |
多肽電泳配套試劑
名稱 |
貨號(hào) |
規(guī)格 |
RTD6120 |
10次 |
|
RTD6101 |
10次(100 μl) |
|
RTD6125 |
10次(50 μl) |
|
RTD6110 |
10次 (50 μl) |
|
RTD6145-01 |
10次 (50 μl) |
|
AC1914-01 |
100ml |
|
AC2914-01 |
100ml |
|
GB010-100ml |
100ml |
|
TP050 |
10×1ml |
|
RTD6202 |
500ml |
|
EA0582-02 |
100 ml |
|
CB010P |
500 ml |
|
AB080 |
250 ml |
|
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 link:http://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 link:https://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
Institution:Institute of Plant Protection, Chinese Academy of Agricultural Sciences
Paper link:https://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)
Institution:College of Biological Science and Biotechnology, Beijing Forestry University
Paper link:https://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
Institution:Institute of Plant Protection, Chinese Academy of Agricultural Sciences
Paper link:https://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(yù)染超低分子量蛋白(1.2-45 kD)
Journal: Plant Cell. Volume 33, Issue 5, May 2021, Pages 1594–1614
Institution:Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing Forestry University
Paper link:https://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(yù)染超低分子量蛋白(1.2-45 kD)
Journal: Analytical Methods. 2021,13, 2804-2811
Institution:Department of Chemistry, Fudan University
Paper link:https://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
Institution:College of Veterinary Medicine, Hunan Agricultural University,
Paper link:https://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
Institution:College of Food Science and Technology, Hebei Normal University of Science and Technology
Paper link:https://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
Institution:Department of Food Processing Technology, Anqing Vocational & Technical College
Paper link:https://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 link:https://pubmed.ncbi.nlm.nih.gov/34996052/
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