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i am working in a lab in japan for 3 weeks, they recently ordered one of your antibodies for me (ab3348 - vamp4). can you please tell me if there is a positive control available for this antibody, ie the epitope used to raise the antibody. we are using it for western blotting but the band apears to be too high a molecular weight, so it would be great to run the positive control. i would really appreciate your help with this, as i have come all the way from edinburgh to japan and really need to know what is going on with this antibody regards |
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ANSWER: |
Thank you for your enquiry regarding ab3348. The positive control for WB is PC3 cell extract which you can purchase from Abcam via our Japanese distributor Cosmobio, catalogue number ab3954. http://www.abcam.com/index.html?datasheet=3954 I would be happy to also look at your protocol and see if there are suggestions I can make to help you. I enclose our WB protocol questionnaire which will help you put your details together, if you can put it to my attention in the additional notes section I will look into this when I receive it. http://www.abcam.com/index.html?section=western&pageconfig=technical&intAbID=3348&mode=questionaire Best of luck with your research in Japan and back home, |
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Anti-VAMP4 antibody (ab3348) at 1 µg/ml +
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 16 kDa
Observed band size : 16 kDa
Additional bands at : 36 kDa,49 kDa,75 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 30 seconds
Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human cervical carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing VAMP4 (ab3348) or without primary antibody (negative control) overnight at 4ºC in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human heart tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing VAMP4 (ab3348) or without primary antibody (negative control) overnight at 4ºC in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1/200 with a rabbit polyclonal antibody recognizing VAMP4 (ab3348) or without primary antibody (negative control) overnight at 4ºC in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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