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ab412 no signal in WB
Using M2A mouse neuroblastoma cell line
Asked on Dec 04 2012
Thank you for your recent telephone call and for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
As requested, I have obtained some more information regaring Western blot protocols using ab412 mono and dimethyl Arginine antibody [7E6] . I hope the following publications in which the antibody has been used in WB will be helpful:
Blifernez O et al. Protein arginine methylation modulates light-harvesting antenna translation in Chlamydomonas reinhardtii. Plant J 65:119-30 (2011).
McBride AE et al. Specific sequences within arginine-glycine-rich domains affect mRNA-binding protein function. Nucleic Acids Res 37:4322-30 (2009).
Gupta P et al. PKCepsilon stimulated arginine methylation of RIP140 for its nuclear-cytoplasmic export in adipocyte differentiation. PLoS ONE 3:e2658 (2008).
As discussed on the telephone, I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.
I would appreciate if you could also provide an image which would help us to assess the results.
Thank you for your time and cooperation. We look forward to receiving the completed questionaire.
Choose: Non-specific band Multiple bands No signal or weak signal High background
Purchase order number
or preferably Abcam order number:
Antibody storage conditions (temperature/reconstitution etc)
Description of the problem (high background, wrong band size, more bands, no band etc.)
Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)
Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)
Amount of protein loaded
Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)
Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)
Detection method (ECL, ECLPlus etc.)
Positive and negative controls used (please specify)
Optimization attempts (problem solving)
How many times have you tried the Western?
Have you run a "No Primary" control?
Do you obtain the same results every time?
e.g. are the background bands always in the same place?
What steps have you altered?
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.
Answered on Dec 04 2012