Anti-CD4 antibody [EPR20122] - BSA and Azide free

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD4 antibody [EPR20122] - BSA and Azide free (AB251480)

This data was developed using ab207755, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Mouse thymocytes labeling CD4 with ab207755 at 1/60 dilution (red), compared with ab172730 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD4 antibody [EPR20122] - BSA and Azide free (AB251480)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207755).

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab207755 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab207755 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 0.2 μg/ml (1/10200)) for 30min on ice. The cells were simultaneously stained with CD3.

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.

• IP

Supplier Data

Immunoprecipitation - Anti-CD4 antibody [EPR20122] - BSA and Azide free (AB251480)

This data was developed using ab207755, the same antibody clone in a different buffer formulation.

CD4 was immunoprecipitated from 0.35 mg of Mouse thymus lysate with ab207755 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207755 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Mouse thymus lysate, 10 μg (Input).
Lane 2 : ab207755 IP in Mouse thymus lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab207755 in Mouse thymus lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-CD4 antibody [EPR20122] (<a href='/en-us/products/primary-antibodies/cd4-antibody-epr20122-ab207755'>ab207755</a>)

Predicted band size: 51 kDa

false

• WB

Supplier Data

Western blot - Anti-CD4 antibody [EPR20122] - BSA and Azide free (AB251480)

This data was developed using ab207755, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CD4 antibody [EPR20122] (<a href='/en-us/products/primary-antibodies/cd4-antibody-epr20122-ab207755'>ab207755</a>) at 1/5000 dilution

All lanes:

Mouse thymus tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 51 kDa

false

Exposure time: 5s