Specific protocols
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To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on. |
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General protocols
Useful resources:Western blotting (WB) protocols:Immunohistochemistry (IHC) / Immunocytochemistry (ICC) protocols:Chromatin Immunoprecipitation (ChIP) protocols:Dot blot protocols:ELISA protocols:ELISPOT protocols: Flow cytometry / FACS protocols:Immunoprecipitation (IP) protocols:
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Western blot - VAMP4 antibody (ab3348)
Anti-VAMP4 antibody (ab3348) at 1 µg/ml + Brain (Human) Tissue Lysate - adult normal tissue (ab29466) at 10 µg
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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody (ab3348)
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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody (ab3348)
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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VAMP4 antibody (ab3348)
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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