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If your product does not perform as described on this datasheet, we will refund or replace your product...
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Thank you soooo much, the antibodys worked and the student finished in time for the presentation. I can not thank you enough. |
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ANSWER: |
I am glad it all worked out for you and the student. |
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LOT NUMBER -- NOT SPECIFIED -- ORDER NUMBER -- NOT SPECIFIED --
DESCRIPTION OF THE PROBLEM Non-specific band
SAMPLE crude extract
PRIMARY ANTIBODY gfap 1:1000 1 hr. 20°C
ANTIBODY STORAGE CONDITIONS -20°C
SAMPLE PREPARATION lysis buffer (tris based)
AMOUNT OF PROTEIN LOADED 15 ug
TRANSFER AND BLOCKING CONDITIONS blocking solution. Milk
SECONDARY ANTIBODY jackson. Rabbit IgG 1:50000
ADDITIONAL NOTES I need to know the epitope for ab7779. Thanks |
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ANSWER: |
Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.
Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab7779. I would also appreciate if you can confirm some further details:
What was the lot number or order number?
What species are your crude extracts from? Have you done a run with an IP first before the WB to isolate and concentrate the GFAP in the sample? Are your extracts from brain tissue?
Do you have a copy of your WB image? Usually .doc, .ppt, .pdf, or .jpg work well. Where are you seeing the multiple bands?
Was your lysis buffer NP-40? Nonidet-P40 (NP-40) buffer: 20 mM Tris HCl pH 8 137 mM NaCl 10% glycerol 1% nonidet P-40 2 mM EDTA
What percentage, time and temperature was the milk blocking step? Have you tried 5% BSA for 1 hr at RT? This is what we tested in-house.
You may want to try the primary at 1:2500 if you're seeing non-specific bands, and incubate this overnight at 4C instead of 1 hr with the 5% blocking buffer.
Since ab7779 is a polyclonal antibody and it was raised against a cow spinal cord prep, it will have multiple binding epitopes along the protein.
Should the suggestions not improve the results, please do let me know.
In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.
I hope this information is helpful, and I thank you for your cooperation. |
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I already purchased the GFAP-antibody (ab7779)from Abcam. I couldn?t find a specification for the protein concentration of this antibody on the datasheet. Can you tell me what?s the antibody concentration in the vial, please? |
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ANSWER: |
Thank you for your enquiry. Ab7779 is supplied as whole antiserum and is therefore unpurified. We therefore cannot determine its IgG concentration as there are other proteins in the solution. We however estimate a concentration of 10-16mg/ml. I hope this estimation will help you. We also have recommended dilutions: IF: 1/1000. IHC-F: 1/500 - 1/1000. IHC-P: 1/500 - 1/1000. IHC-Fr: 1/500 - 1/1000. |
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