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Tissue, cells or virus corresponding to Human CD81. Aggregated OCI-LY8 cells.
Ab35026 is a potent CD81 reagent, induces homotypic adhesion and has powerful anti proliferative effects. Endotoxin Level: <0.01 Eu/ug (low endotoxin grade).
Our Abpromise guarantee covers the use of ab35026 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 10µl of the suggested working dilution to label 106 cells in 100µl.
|IHC-P||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Use under non reducing condition. Predicted molecular weight: 26 kDa.
|Functional Studies||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
Overlay histogram showing HAP1 wildtype (green line) and HAP1-CD81 knockout cells (red line) stained with ab35026. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab35026, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) presorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.
A mouse IgG1 isotype control antibody (ab170190) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CD81 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
ab35026 staining CD81 in SH-SY5Y were transfected with cDNA encoding CD81 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 3% BSA for 2 hours at room temperature. Samples were incubated with primary antibody (1/100 in PBS + 3% BSA) for 2 hours. An Alexa Fluor® 488-conjugated goat anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.
Flow cytometry analysis of Human peripheral blood lymphocytes, staining CD81 with ab35026 at 1/10 dilution.
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