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Our Abpromise guarantee covers the use of ab97046 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. (Primary)|
|WB||1/2000 - 1/20000. (Colorimetric). (Chemiluminescent 1/10000 - 1/50000).|
IHC image of beta actin 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 ab8224, 3µg/ml overnight at +4°C. An HRP-conjugated secondary (ab97046, 1/500 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-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml
Lanes 1-6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Lanes 1 - 2 : Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab97046) at 1/2000 dilution
Lanes 3 - 4 : Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab97046) at 1/10000 dilution
Lanes 5 - 6 : Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab97046) at 1/20000 dilution
Developed using the
Sample: Human Cell lysate - whole cell (human glioma cells), Loading amount: 20 µg, Specification: human glioma cells
Blocking step: Milk as blocking agent for 1 hour, Concentration: 5 %, Temperature: 25°C
Other product details: 1st Ab Dilution 1/5000 anti-actin Ab, Incubation time: 16 hours, Temperature: 4°C
Secondary antibody: ab97046 used at 1:20,000
Bands: Specific: 45 kDa
Notes: primary Ab (anti-actin) was used as a loading control in 6 human glioma cell lines
This image is courtesy of an anonymous Abreview