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非變性蛋白電泳Marker轉膜問(wèn)題

http://www.chrismariegreen.com/news_detail.aspx?NewsID=248

寬分子量非變性電泳蛋白質(zhì)Marker（21-880 kD）

Broad MW Native Electrophoresis Protein Marker（21-880 kD）

●產(chǎn)品編號及規格：

RTD6144   20次 100 μl

●儲存及運輸條件：

-20℃保存，濕冰運輸，有效期為12個(gè)月。

● 產(chǎn)品簡(jiǎn)介：

    本產(chǎn)品有5種蛋白組成，分子量范圍為21-880 kD，經(jīng)過(guò)非變性電泳后，用考馬斯亮藍染色后可以得到5條主帶。

● 使用說(shuō)明：

1. 取出產(chǎn)品后，常溫融化后，徹底混勻，上樣電泳。

注：上樣量根據膠的厚度和梳子的寬度確定。一般說(shuō)來(lái)，1.0厚度10齒梳子加樣孔上樣5 μl，1.0厚度15齒梳子加樣孔上樣量2.5 μl，其他規格梳子請適當調整上樣量。

2. 該產(chǎn)品不含蛋白凝膠制備試劑，您可以選擇非變性PAGE凝膠制備及電泳試劑盒（貨號：RTD6130）或者蛋白預制膠（貨號RTD6118）。如自備試劑，經(jīng)典Laemmli系統中把制膠溶液、電泳緩沖液和上樣緩沖液中的SDS和還原劑（DTT或β-巰基乙醇）全部去除即可進(jìn)行非變性電泳。

3. 電泳條件：

建議使用10%非變性膠或者4-20%梯度非變性膠（貨號：RTD6118-0420）進(jìn)行電泳。

4. 電泳結束后，考馬斯亮藍染色，觀(guān)察結果。

注：使用銀染染色時(shí)，由于靈敏度高于考馬斯亮藍染色方法，需要適當降低Marker上樣量，一般稀釋50倍后上樣。

注意：

1. 本蛋白Marker不適用于變性蛋白電泳（SDS-PAGE），因為在SDS存在下，含有多個(gè)亞單位的蛋白會(huì )不同程度解聚。

2. 在非變性條件下，蛋白的遷移與蛋白的電荷、蛋白形狀以及蛋白分子量都有關(guān)。因此，在一種凝膠濃度下使用本Marker不能精確估計出目的蛋白的分子量。非變性電泳中，蛋白分子量的確定應該是在不同凝膠濃度下，確定出蛋白的Rf值，繪制出凝膠濃度對Rf的曲線(xiàn)從而判定蛋白的分子量。

● 實(shí)驗示例：

1 4-20% Tris-甘氨酸 非變性膠電泳：

    4-20% RealPAGE Precast Gel
1×TG 200V 50-19mA  47min

2. 4-16% Blue Native：

3： 4-15% Tris-甘氨酸非變性：

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 garvieae isolated 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/