Home About us Editorial board Search Ahead of print Current issue Archives Submit article Instructions Subscribe Contacts Login 
Visit old site
Home Print this page Email this page Small font size Default font size Increase font size
Users Online: 463

 Table of Contents  
Year : 2014  |  Volume : 6  |  Issue : 3  |  Page : 160

Western blot: Technique, theory and trouble shooting

Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada

Date of Web Publication11-Mar-2014

Correspondence Address:
Ping-Chang Yang
Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/1947-2714.128482

Rights and Permissions

How to cite this article:
Liu ZQ, Mahmood T, Yang PC. Western blot: Technique, theory and trouble shooting. North Am J Med Sci 2014;6:160

How to cite this URL:
Liu ZQ, Mahmood T, Yang PC. Western blot: Technique, theory and trouble shooting. North Am J Med Sci [serial online] 2014 [cited 2020 Sep 19];6:160. Available from: http://www.najms.org/text.asp?2014/6/3/160/128482

Dear Editor,

We appreciate the comments [1] regarding our published article "Western blot: Technique, theory, and trouble-shooting". [2] For transferring of proteins to a membrane, the method of "wet transfer" in Western blotting is routinely utilized by laboratories and guided by the protocols online. [3] Heat may be produced during the transfer; according to the recommended protocols, placing the transfer apparatus on ice or placing it in the cold room is necessary. It should be considered that the purpose of 'Figure 7' in our article [2] is to show how to cool down the environmental temperature to 4°C by placing the apparatus on ice, which cannot be misunderstood as submerging the electroporator or the power leads in the water bath. In addition, we agree with the proposal that safe laboratory practice should always be observed to avoid chemical or electrical hazards in operating the Western blot transferring systems.

  References Top

1.Gibbons J. Western blot: Protein transfer overview. N Am J Med Sci 2014;6:158-9.  Back to cited text no. 1
2.Mahmood T, Yang PC. Western blot: Technique, theory, and trouble shooting. N Am J Med Sci 2012;4:429-34.  Back to cited text no. 2
3.Protocol Online. (Accessed December 25, 2013, at www.protocolonline.org/prot/Molecular_Biology/Protein/Western_Blotting ).  Back to cited text no. 3

This article has been cited by
1 Vorinostat is genotoxic and epigenotoxic in the mouse bone marrow cells at the human equivalent doses
Sabry M. Attia,Mohamed K. Al-Khalifa,Mohammed A. Al-Hamamah,Moureq R. Alotaibi,Mohamed S.M. Attia,Sheikh F. Ahmad,Mushtaq A. Ansari,Ahmed Nadeem,Saleh A. Bakheet
Toxicology. 2020; 441: 152507
[Pubmed] | [DOI]
2 Effects of Salt Stimulation on Lunasin Accumulation and Activity during Soybean Germination
Weiyi Zhang,Yuqiong Hao,Cong Teng,Xin Fan,Xiushi Yang,Mengjie Liu,Guixing Ren,Congping Tan
Foods. 2020; 9(2): 118
[Pubmed] | [DOI]
3 Limonene, a food additive, and its active metabolite perillyl alcohol improve regeneration and attenuate neuropathic pain after peripheral nerve injury: Evidence for IL-1ß, TNF-a, GAP, NGF and ERK involvement
Heitor G. Araújo-Filho,Erik W.M. Pereira,Luana Heimfarth,Brenda Souza Monteiro,Fabiolla Rocha Santos Passos,Pollyana Siqueira-Lima,Sathiyabama R. Gandhi,Márcio Roberto Viana dos Santos,Jackson Roberto Guedes da Silva Almeida,Laurent Picot,Raphaël Grougnet,Ray S. Almeida,Henrique Douglas Melo Coutinho,Lucindo J. Quintans-Júnior,Natália Martins,Jullyana S.S. Quintans
International Immunopharmacology. 2020; 86: 106766
[Pubmed] | [DOI]
4 Effect of Flexible and Rigid Linkers on Biological Activity of Recombinant Tetramer Variants of S3 Antimicrobial Peptide
Sakineh Baghbeheshti,Shahin Hadadian,Akram Eidi,Leila Pishkar,Hamzeh Rahimi
International Journal of Peptide Research and Therapeutics. 2020;
[Pubmed] | [DOI]
5 Preeclampsia serum increases CAV1 expression and cell permeability of human renal glomerular endothelial cells via down-regulating miR-199a-5p, miR-199b-5p, miR-204
Fei Du,Ye Zhang,Qinyang Xu,Yincheng Teng,Minfang Tao,Alex F. Chen,Rongzhen Jiang
Placenta. 2020; 99: 141
[Pubmed] | [DOI]
6 The Three Lipocalins of Egg-White: Only Ex-FABP Inhibits Siderophore-Dependent Iron Sequestration by Salmonella Enteritidis
Louis Alex Julien,Clémence Fau,Florence Baron,Sylvie Bonnassie,Catherine Guérin-Dubiard,Françoise Nau,Michel Gautier,Kimon Andreas Karatzas,Sophie Jan,Simon Colin Andrews
Frontiers in Microbiology. 2020; 11
[Pubmed] | [DOI]
7 Peg-Enhanced Behavioral Recovery After Sciatic Nerve Transection and Either Suturing Or Sleeve Conduit Deployment in Rats
Xin Zhao,Bo Li,Xiangchen Guan,Guiyin Sun,Mingzhe Zhang,Weihua Zhang,Junfeng Xu,Xiaoping Ren
Journal of Investigative Surgery. 2019; : 1
[Pubmed] | [DOI]
8 The IL-37–Mex3B–Toll-like receptor 3 axis in epithelial cells in patients with eosinophilic chronic rhinosinusitis with nasal polyps
Jin-Xin Liu,Bo Liao,Qi-Hong Yu,Hai Wang,Yi-Bo Liu,Cui-Lian Guo,Zhi-Chao Wang,Zhi-Yong Li,Zhe-Zheng Wang,Jian-Wen Ruan,Li Pan,Yin Yao,Cai-Ling Chen,Heng Wang,Yuxia Liang,Guohua Zhen,Zheng Liu
Journal of Allergy and Clinical Immunology. 2019;
[Pubmed] | [DOI]
9 Structure-based design and synthesis of new 4-methylcoumarin-based lignans as pro-inflammatory cytokines (TNF-a, IL-6 and IL-1ß) inhibitors
S Santhosh Kumar,Ahil Sajeli Begum,Kirti Hira,Sarfaraj Niazi,B.R. Prashantha Kumar,Hiroshi Araya,Yoshinori Fujimoto
Bioorganic Chemistry. 2019; : 102991
[Pubmed] | [DOI]
10 The impact of concurrent training and antioxidant supplementation on the factors associated with the ocular lens opacity in diabetic rats
Mousa Amirahmadi,Mohsen Salesi,Reza Yousefi,Farhad Daryanosh,Javad Nemati,Boris I. Kurganov
Archives of Physiology and Biochemistry. 2019; : 1
[Pubmed] | [DOI]
11 Genetic polymorphism in core promoter sequence of ACTRIIB gene and association analysis with growth traits in chicken
Pothanaboyina Guru Vishnu,Tarun Kumar Bhattacharya,Bharat Bhushan,Chandan Paswan,Athe Rajendra Prasad,Devara Divya
Reproduction in Domestic Animals. 2019;
[Pubmed] | [DOI]
12 Variable impact of the hemagglutinin polybasic cleavage site on virulence and pathogenesis of avian influenza H7N7 virus in chickens, turkeys and ducks
David Scheibner,Reiner Ulrich,Olanrewaju I. Fatola,Annika Graaf,Marcel Gischke,Ahmed H. Salaheldin,Timm C. Harder,Jutta Veits,Thomas C. Mettenleiter,Elsayed M. Abdelwhab
Scientific Reports. 2019; 9(1)
[Pubmed] | [DOI]
13 Niclosamide reverses adipocyte induced epithelial-mesenchymal transition in breast cancer cells via suppression of the interleukin-6/STAT3 signalling axis
Jones Gyamfi,Yun-Hee Lee,Byung Soh Min,Junjeong Choi
Scientific Reports. 2019; 9(1)
[Pubmed] | [DOI]
14 Can We Use 2,3,5-Triphenyltetrazolium Chloride-Stained Brain Slices for Other Purposes? The Application of Western Blotting
Sonia Sanchez-Bezanilla,Michael Nilsson,Frederick R. Walker,Lin Kooi Ong
Frontiers in Molecular Neuroscience. 2019; 12
[Pubmed] | [DOI]
15 Mechanism of SPRY1 methylation regulating natural aging of skin epidermal cells
E Yang,Zhao Juan,Zhang Hengshu
Journal of Cosmetic Dermatology. 2019;
[Pubmed] | [DOI]
16 LncRNA SNHG16 induces the SREBP2 to promote lipogenesis and enhance the progression of pancreatic cancer
Yi Yu,Jia-Tian Dong,Bing He,Yu-Feng Zou,Xue-Song Li,Chen-Hui Xi,Yuan Yu
Future Oncology. 2019;
[Pubmed] | [DOI]
17 CRISPR/Cas9-Mediated BRCA1 Knockdown Adipose Stem Cells Promote Breast Cancer Progression
Ruya Zhao,Rayan Kaakati,Xinjian Liu,Lingfan Xu,Andrew K. Lee,Robin Bachelder,Chuan-Yuan Li,Scott T. Hollenbeck
Plastic and Reconstructive Surgery. 2019; 143(3): 747
[Pubmed] | [DOI]
18 Effects of Eclipta prostrata on gut microbiota of SAMP6 mice with osteoporosis
Xin Zhao,Juqing Ai,Haoping Mao,Xiumei Gao
Journal of Medical Microbiology. 2019; 68(3): 402
[Pubmed] | [DOI]
19 Combretastatin A4 nanodrug combined plerixafor for inhibiting tumor growth and metastasis simultaneously
Jian Jiang,Na Shen,Wantong Song,Haiyang Yu,Kazuo Sakurai,Zhaohui Tang,Gao Li
Biomaterials Science. 2019;
[Pubmed] | [DOI]
20 Lobaplatin inhibits prostate cancer progression in part by impairing AR and ERG signal
Lei Chen,Hongwen Cao,Chao Yu,Yigeng Feng
Fundamental & Clinical Pharmacology. 2018;
[Pubmed] | [DOI]
21 Defining the molecular signatures of human right heart failure
Jordan L. Williams,Omer Cavus,Emefah C. Loccoh,Sara Adelman,John C. Daugherty,Sakima A. Smith,Benjamin Canan,Paul M.L. Janssen,Sara Koenig,Crystal F. Kline,Peter J. Mohler,Elisa A. Bradley
Life Sciences. 2018; 196: 118
[Pubmed] | [DOI]
22 Expression of Bioactive Lunasin Peptide in Transgenic Rice Grains for the Application in Functional Food
Guixing Ren,Yuqiong Hao,Yingying Zhu,Zhenxing Shi,Gang Zhao
Molecules. 2018; 23(9): 2373
[Pubmed] | [DOI]
23 Photodynamic inactivation of non-enveloped RNA viruses
Hussaini Majiya,Oluwapelumi O. Adeyemi,Morgan Herod,Nicola J. Stonehouse,Paul Millner
Journal of Photochemistry and Photobiology B: Biology. 2018; 189: 87
[Pubmed] | [DOI]
24 Hollow Nanoneedle-Electroporation System To Extract Intracellular Protein Repetitively and Nondestructively
Gen He,Chengduan Yang,Tian Hang,Di Liu,Hui-Jiuan Chen,Ai-hua Zhang,Dian Lin,Jiangming Wu,Bo-ru Yang,Xi Xie
ACS Sensors. 2018;
[Pubmed] | [DOI]
25 Polygodial analog induces apoptosis in LNCaP prostate cancer cells
Subramanyam Dasari,Angela Lincy Prem Antony Samy,Parnal Narvekar,Venkata Satish Dontaraju,Ramesh Dasari,Alexander Kornienko,Gnanasekar Munirathinam
European Journal of Pharmacology. 2018;
[Pubmed] | [DOI]
26 Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer
Hui-Yu Chuang,Jeng-Kae Jiang,Muh-Hwa Yang,Hsei-Wei Wang,Ming-Chun Li,Chan-Yen Tsai,Yau-Yun Jhang,Jason C. Huang
Oncotarget. 2017; 8(13): 21266
[Pubmed] | [DOI]
27 Interferon-?-induced insufficient autophagy contributes to p62-dependent apoptosis of epithelial cells in chronic rhinosinusitis with nasal polyps
B-F. Wang,P-P. Cao,Z-C. Wang,Z-Y. Li,Z-Z. Wang,J. Ma,B. Liao,Y-K. Deng,X-B. Long,K. Xu,H. Wang,H. Wang,M. Zeng,X. Lu,Z. Liu
Allergy. 2017;
[Pubmed] | [DOI]
28 Evaluation of Nanobody Conjugates and Protein Fusions as Bioanalytical Reagents
Virginia J. Bruce,Brian R. McNaughton
Analytical Chemistry. 2017; 89(7): 3819
[Pubmed] | [DOI]
29 Establishment of a human hepatocellular cell line capable of maintaining long-term replication of hepatitis B virus
Wan-Ling Yao,Sotaro Ikeda,Yuta Tsukamoto,Keiko Shindo,Yukie Otakaki,Mian Qin,Yoshikazu Iwasawa,Fumihiko Takeuchi,Yuki Kaname,Yu-Chi Chou,Chungming Chang,Koichi Watashi,Takaji Wakita,Takeshi Noda,Hiroki Kato,Takashi Fujita
International Immunology. 2017; 29(3): 109
[Pubmed] | [DOI]
30 Niclosamide induces apoptosis through mitochondrial intrinsic pathway and inhibits migration and invasion in human thyroid cancer in vitro
Kai Yu,Tingting Wang,Yujue Li,Chun Wang,Xia Wang,Mei Zhang,Yongmei Xie,Shuangqing Li,Zhenmei An,Tinghong Ye
Biomedicine & Pharmacotherapy. 2017; 92: 403
[Pubmed] | [DOI]
31 Celecoxib Enhances the Efficacy of Low-Dose Antibiotic Treatment against Polymicrobial Sepsis in Mice and Clinical Isolates of ESKAPE Pathogens
Madhavi Annamanedi,Gajapati Y. N. Varma,K. Anuradha,Arunasree M. Kalle
Frontiers in Microbiology. 2017; 8
[Pubmed] | [DOI]
32 Increasing Type 1 Poliovirus Capsid Stability by Thermal Selection
Oluwapelumi O. Adeyemi,Clare Nicol,Nicola J. Stonehouse,David J. Rowlands,Terence S. Dermody
Journal of Virology. 2017; 91(4)
[Pubmed] | [DOI]
33 Multiscalar cellular automaton simulates in-vivo tumour-stroma patterns calibrated from in-vitro assay data
J. A. Delgado-SanMartin,J. I. Hare,E. J. Davies,J. W. T. Yates
BMC Medical Informatics and Decision Making. 2017; 17(1)
[Pubmed] | [DOI]
34 A prosurvival and proangiogenic stem cell delivery system to promote ischemic limb regeneration
Yanyi Xu,Minghuan Fu,Zhihong Li,Zhaobo Fan,Xiaofei Li,Ying Liu,Peter M. Anderson,Xiaoyun Xie,Zhenguo Liu,Jianjun Guan
Acta Biomaterialia. 2016; 31: 99
[Pubmed] | [DOI]
35 Protein stability: a crystallographeræs perspective
Marc C. Deller,Leopold Kong,Bernhard Rupp
Acta Crystallographica Section F Structural Biology Communications. 2016; 72(2): 72
[Pubmed] | [DOI]
36 Analysis of protein phosphorylation in nerve terminal reveals extensive changes in active zone proteins upon exocytosis
Mahdokht Kohansal-Nodehi,John JE Chua,Henning Urlaub,Reinhard Jahn,Dominika Czernik
eLife. 2016; 5
[Pubmed] | [DOI]
37 UBC9-Mediated Sumoylation Favorably Impacts Cardiac Function in Compromised Hearts
Manish K. Gupta,Patrick M. McLendon,James Gulick,Jeanne James,Kamel Khalili,Jeffrey Robbins
Circulation Research. 2016; 118(12): 1894
[Pubmed] | [DOI]


Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

  In this article

 Article Access Statistics
    PDF Downloaded4031    
    Comments [Add]    
    Cited by others 37    

Recommend this journal