Anti-CD80 antibody [EPR22183] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD80 antibody [EPR22183] - BSA and Azide free (AB238648)

Immunofluorescent analysis of 100% methanol-fixed Raji (human Burkitt's lymphoma cell line) and HUVEC (human umbilical vein endothelial cell line) cells labeling CD80 with ab225674 at 1/1000 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 in Raji cell line is observed. 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).

PBS only control : Used PBS instead of primary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

Negative control : HUVEC cells (PMID : 15064090).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab225674).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD80 antibody [EPR22183] - BSA and Azide free (AB238648)

Flow cytometric analysis of HUVEC (human umbilical vein endothelial cell line) (left panel) cells and Raji (human Burkitt's lymphoma cell line)(right panel) cells labeling CD80  with ab225674 at 1/400 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

Negative control : HUVEC (PMID : 15064090).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab225674).

• Flow Cyt

Lab

Flow Cytometry - Anti-CD80 antibody [EPR22183] - BSA and Azide free (AB238648)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab225674).

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) (top) or PBMCs treated with CD3/CD28 activation (bottom), with ab225674 (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 interactions, followed by the antibody ab225674 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ in 100 µl at 5.0 μg/ml (1/400)) for 30 min at 22°C . Cells were simultaneously stained with CD4

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/2000 for 30min 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 live cells.

• IP

Unknown

Immunoprecipitation - Anti-CD80 antibody [EPR22183] - BSA and Azide free (AB238648)

CD80 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab225674 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab225674 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Raji whole cell lysate 10 μg (Input).
Lane 2 : ab225674 IP in Raji whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab225674 in Raji whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 1 second.

This blot was developed using a higher sensitivity ECL substrate.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab225674).

All lanes:

Immunoprecipitation - Anti-CD80 antibody [EPR22183] (<a href='/en-us/products/primary-antibodies/cd80-antibody-epr22183-ab225674'>ab225674</a>)

Predicted band size: 33 kDa

Observed band size: 60 kDa

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