Anti-CD40 antibody [EPR18005-35]

• Flow Cyt

Lab

Flow Cytometry - Anti-CD40 antibody [EPR18005-35] (AB188181)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab188181 (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 ab188181 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 5.0 μg/ml (1/388)) 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR18005-35] (AB188181)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A20 (mouse reticulum sarcoma cell line) cells labeling CD40 with ab188181 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on A20 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD40 antibody [EPR18005-35] (AB188181)

Flow cytometric analysis of A20 (mouse reticulum sarcoma cell line) cell line labeling CD40 with ab188181 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-CD40 antibody [EPR18005-35] (AB188181)

Blocking/Dilution : 5% NFDM/TBST.

Exposure : 15 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 23404288 and PMID : 16709811).

All lanes:

Western blot - Anti-CD40 antibody [EPR18005-35] (ab188181) at 1/1000 dilution

All lanes:

A20 (mouse reticulum sarcoma cell line) whole cell 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 1/20000 dilution

Predicted band size: 30 kDa

Observed band size: 32-42 kDa

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