Anti-CD79a antibody [EPR3619]

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD79a antibody [EPR3619] (AB133483)

Flow cytometric analysis of permeabilised Ramos cells labelled with ab133483 at 1/100 dilution (red) or negative rabbit IgG (green).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD79a antibody [EPR3619] (AB133483)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab133483 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 mins on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab133483 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ in 100 µl at 0.008 μg/ml (1/62,5000 dilution) for 30 mins at 22°C . The cells were simultaneously stained with CD19.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30 mins at 22°C

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on CD19+.

• WB

Unknown

Western blot - Anti-CD79a antibody [EPR3619] (AB133483)

All lanes:

Western blot - Anti-CD79a antibody [EPR3619] (ab133483) at 1/1000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

Ramos cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 25 kDa

false

• WB

Lab

Western blot - Anti-CD79a antibody [EPR3619] (AB133483)

False colour image of Western blot : Anti-CD79a antibody [EPR3619] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133483 was shown to bind specifically to CD79a. A band was observed at 50 kDa in wild-type Raji cell lysates with no signal observed at this size in CD79A knockout cell line ab274911 (knockout cell lysate ab281361). To generate this image, wild-type and CD79A knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-CD79a antibody [EPR3619] (ab133483) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

CD79A knockout Raji cell lysate at 20 µg

Lane 3:

Daudi cell lysate at 20 µg

Secondary

Lanes 1 - 3:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 3:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 50 kDa

false