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Our Abpromise guarantee covers the use of ab97100 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 concentration.|
|IHC-P||1/200 - 1/5000.|
|ELISA||1/10000 - 1/100000.|
|WB||1/2000 - 1/20000. Colorimetric: 1/5000 - 1/30000; Chemiluminescent: 1/10000 - 1/50000|
IHC image of beta Actin staining in Human normal 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 ab8229, 10µg/ml overnight at +4°C. An HRP-conjugated secondary (Ab97100, 1/100 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
The inset negative control image is secondary-only at 1/500 dilution.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
All lanes :
Anti-beta Actin antibody (ab8229) at 1 µg/ml and HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg per lane.
Lanes 1 - 2 : Rabbit polyclonal Secondary Antibody to Goat IgG - H&L (HRP) (ab97100) at 1/5000 dilution
Lanes 3 - 4 : Rabbit polyclonal Secondary Antibody to Goat IgG - H&L (HRP) (ab97100) at 1/10000 dilution
Lanes 5 - 6 : Rabbit polyclonal Secondary Antibody to Goat IgG - H&L (HRP) (ab97100) at 1/20000 dilution
Developed using the ECL technique
Performed under reducing conditions.
Exposure time : 8 minutes