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Our Abpromise guarantee covers the use of ab97057 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use at an assay dependent dilution.|
|IHC-P||1/200 - 1/5000.|
|ELISA||1/10000 - 1/100000. (as primary)|
|WB||1/2000 - 1/20000. Colorimetric: Use 1/5000-1/30000, Chemiluminescent: use 1/10000-1/50000|
IHC image of alpha tubulin staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab64332, 5µg/ml overnight at +4°C. An HRP-conjugated secondary (ab97057, 1/200 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
The inset negative control image is taken from an identical assay without primary antibody.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
All lanes : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/ml
Lanes 1-6: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Lanes 1 - 2 : Goat polyclonal Secondary Antibody to Rat IgG - H&L (HRP) (ab97057) at 1/2000 dilution
Lanes 3 - 4 : Goat polyclonal Secondary Antibody to Rat IgG - H&L (HRP) (ab97057) at 1/10000 dilution
Lanes 5 - 6 : Goat polyclonal Secondary Antibody to Rat IgG - H&L (HRP) (ab97057) at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size : 50 kDa
Observed band size : 52 kDa
Additional bands at : 85 kDa. We are unsure as to the identity of these extra bands.