Anti-CD79b antibody [EPR6860]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CD79b antibody [EPR6860] (AB134103)

Immunofluorescence analysis of CD79b in Ramos cells, using ab134103 antibody at a dilution of 1/250.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD79b antibody [EPR6860] (AB134103)

Flow Cytometry analysis of Raji cells labeling CD79b with ab134103 at 1/20 dilution (red). A rabbit IgG was used as negative control (green).

• Flow Cyt

Lab

Flow Cytometry - Anti-CD79b antibody [EPR6860] (AB134103)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab134103 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). PBMCs were incubated for 30 min 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 ab134103 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 1.0 μg/ml (1/2300)) for 30min on ice. 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 for 30min on ice

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

• WB

Lab

Western blot - Anti-CD79b antibody [EPR6860] (AB134103)

False colour image of Western blot : Anti-CD79b antibody [EPR6860] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134103 was shown to bind specifically to CD79b. A band was observed at 39 kDa in wild-type Raji cell lysates with no signal observed at this size in CD79B CRISPR-Cas9 edited cell line ab273839 (CRISPR-Cas9 edited cell lysate ab273793). The band observed in the CRISPR-Cas9 edited lysate lane below 39 kDa is likely to represent a truncated form of CD79b. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and CD79B CRISPR-Cas9 edited Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-CD79b antibody [EPR6860] (ab134103) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

CD79B CRISPR-Cas9 edited Raji cell lysate at 20 µg

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

HAP1 cell lysate at 20 µg

Observed band size: 39 kDa

false

• WB

Unknown

Western blot - Anti-CD79b antibody [EPR6860] (AB134103)

All lanes:

Western blot - Anti-CD79b antibody [EPR6860] (ab134103) at 1/1000 dilution

Lane 1:

Daudi cell lysate at 10 µg

Lane 2:

Namalwa cell lysate at 10 µg

Lane 3:

Ramos cell lysate at 10 µg

Lane 4:

Raji cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 26 kDa

Observed band size: 35-40 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CD79b antibody [EPR6860] (AB134103)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.