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- Proteins and Peptides
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If your product does not perform as described on this datasheet, we will refund or replace your product...
Read our guarantee »This product is covered by the Abpromise guarantee. Our scientific support team are available to answer any questions or queries - fill out an inquiry form for ab2910 for help.
Alternatively, you can search the previous enquiries about this product to see if your query has already been answered.
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Our customer purchased ab2910(lot#GR54402-1), ab2911(lot#GR11226-5) and would like to know KD values of these antibodies, respectively. |
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ANSWER: |
Thank you for your enquiry. |
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thank you very much for your reply. I would like once more to disturb you and ask the following question; quite often you are indicating that antigen was a synthetic peptide. I am working mainly with chick brain and quite often it is impossible to understand if the antibody is reactive with chicks. I am interested if any of your caveolin antibodies (e.g. 93952, 2910 or any other) is reactive against chicken proteins and the same question with antibody 115822 (Anti-eIF2 alpha). |
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ANSWER: |
Thank you very much for your interest in Abcam antibodies. |
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mouse neuroblastoma cells problem: high background, no specific staining, precipitate seen Ab: 1/250 and 1/500 block: 10% normal goat serum + 6% BSA secondary: AlexaFluor, works well, did no-primary crtl (no signal) |
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ANSWER: |
Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.
I am sorry these 2 products did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued for each a free of charge replacement with the order number xxx.
To check the status of the order please contact our Customer Service team and reference this number.
Please note that these free of charge replacement vials are also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.
I wish you the best of luck with your research. |
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BATCH NUMBER 134228 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM X-ray film is completely blank SAMPLE EL4 and Ramos cells were subjected to lipid raft extraction using an extraction buffer (1% Triton X-100, 25mM Tris-HCl, 150 mM NaCl, 5mM EDTA and protease cocktail) and further subjected to a sucrose density gradient PRIMARY ANTIBODY Abcam rabbit polyclonal to caveolin-1 (ab2910). Blocking buffer was removed by rinsing blot with PBS-0.5% Tween-20. A 1:1000 dilution was used. In the first attempt the blot was incubated in primary ab for 1hr which gave completely blank film (this occurred on two separate occasions). Most recent attempt the blot was incubated with primary ab for 18hrs at 4C DETECTION METHOD Amersham ECL ANTIBODY STORAGE CONDITIONS Antibody aliquots were stored at -20 C SAMPLE PREPARATION sucrose gradient fractions were added to the appropriate amount of NuPAGE LDS Sample buffer, heated at approximately 100C for 10 min prior to being loaded ELECTROPHORESIS/GEL CONDITIONS Non-Reducing conditions. Samples loaded on a 10% Bis-Tris NuPAGE gel and run with 1x MOPS-SDS running buffer at 200V for about 1hr TRANSFER AND BLOCKING CONDITIONS Protein was transferred to nitrocellulose membrane with NuPAGE transfer buffer (500mM Bicine, 500mM Bis-Tris, 20.5mM EDTA, 1mM Chlorobutanol) at 30V for 1hr 30min. Blot was subsquently blocked with 5% Skim milk PBS-0.5% Tween-20 for 1hr30min SECONDARY ANTIBODY Primary ab was removed by 3 x 5 min washes with PBS-0.5% Tween-20. A 1:4000 dilution of secondary Ab ([another company's] ECL Anti-rabbit IgG HRP linked, from donkey) was used and incubated for 1 hr. This secondary antibody was used with the same samples and different primary ab and worked perfectly fine HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Altered the duration of the primary ab incubation (as detailed above) ADDITIONAL NOTES We have used nearly the complete stock of ab2910 ordered and still unable to get even the slightest band on the x-ray film and would really appreciate some feedback as soon as possible. As mentioned the samples used in this western were prepped and loaded in the exact same fashion for blots using other primary antibodies and they all worked.
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ANSWER: |
Thank you for taking the time to contact me regarding the problems that you have experienced using this product in Western blotting. Since the target interacts directly with G-protein alpha subunits and can functionally regulate their activity I would advise that the Western blot is carried out in reducing conditions and not non reducing conditions. The aim of reducing conditions being to disrupt disulphide bonds and intermolecular interactions and allow separation based on charge and molecule size. Membrane proteins are especially prone to aggregation which may lead to anomalous protein electrophoretic mobility. I would advise that you alter the incubation period from 100 degrees for 10 mins to 37 degrees for one hour (in the presence of a reducing agent such as DTT). The use of a positive control would also be advantageous to discount any effect on the target by the lipid extraction procedure. The suggested positive control would be human heart, spleen or lung. I hope that this information is useful to you, if you are still experiencing problems utilising this product following the above suggestions then please do not hesitate to get back in touch with me. Good luck with your research.
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I want to use this product to do Immunofluorescence on Bovine Aortic Endothelia Cells(BAEC). But I want to check the caveolin 1 on the membrane but not inside the cells. I want to check the effect of some chemicals on the distribution of caveolin 1 ON the MEMBRANE. Is this product suitable for this purpose?
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ANSWER: |
Thank you for your enquiry. Unfortunately Caveolin 1 antibody - Caveolae Marker (ab2910) has not been tested for use by immunocytochemistry or immunohistochemistry. However, following a brief search of our catalogue I have determined that ab27514 Caveolin 1 antibody matches your requirements. It is rabbit polyclonal antibody that has been shown to work by immunofluorescence (immunocytochemistry) and has several favourable reviews by our customers. You may wish to examine the datasheets and the "Abreview" left by our customers. I hope this information helps, please do not hesitate to contact us if you need any more advice or information. |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
