Anti-CD11a antibody [EPR26928-111A]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD11a antibody [EPR26928-111A] (AB307658)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling CD11a with ab307658 at 1/50 (9.36 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous staining in THP-1 cell line.Negative control : HeLa (PMID : 12036881).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt

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Flow Cytometry - Anti-CD11a antibody [EPR26928-111A] (AB307658)

Flow cytometric analysis of THP-1 (human monocytic leukemia monocyte) cells labelling CD11a with ab307658 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

• Flow Cyt

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Flow Cytometry - Anti-CD11a antibody [EPR26928-111A] (AB307658)

Flow cytometric analysis of HeLa (human cervix adenocarcinoma epithelial cell, Left) / Jurkat (human T cell leukemia T lymphocyte, Right) cells labelling CD11a with ab307658 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Negative control : Hela (PMID : 12036881).

• Flow Cyt

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Flow Cytometry - Anti-CD11a antibody [EPR26928-111A] (AB307658)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling CD11a with ab307658 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Cells were stained with rabbit IgG or ab307658. Then stained with anti-CD3 conjugated to Alexa Fluor® 647.

• IP

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Immunoprecipitation - Anti-CD11a antibody [EPR26928-111A] (AB307658)

CD11a was immunoprecipitated from 0.35 mg THP-1 whole cell lysate with ab307658 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307658 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : THP-1 (human monocytic leukemia monocyte) whole cell lysate Lane 2 : ab307658 IP in THP-1 whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307658 in THP-1 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-CD11a antibody [EPR26928-111A] (ab307658) at 1/30 dilution

All lanes:

THP-1 (human monocytic leukemia monocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 15s

• WB

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Western blot - Anti-CD11a antibody [EPR26928-111A] (AB307658)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : K562 (PMID : 7497527), Hela (PMID : 12036881). The observed Mw of 180KDa is higher than the predicted Mw may due to glycosylation. Exposure time : 37 seconds

All lanes:

Western blot - Anti-CD11a antibody [EPR26928-111A] (ab307658) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 180 kDa

false

Exposure time: 37s