Anti-CD11a antibody [EP1285Y]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11a antibody [EP1285Y] (AB52895)

Immunohistochemical analysis of paraffin-embedded Human tonsil labeling CD11a with ab52895 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab79051) at 1/500 dilution. Membrane/cytoplasm staining on lymphocytes of human tonsil is observed. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt

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Flow Cytometry - Anti-CD11a antibody [EP1285Y] (AB52895)

Flow cytometry analysis of 2% paraformaldehyde fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling CD11a with ab52895 at 1/50 dilution (red line). Secondary antibody used is a goat anti rabbit IgG (FITC) at 1/150 dilution. The isotype control is rabbit monoclonal IgG (black line). The unlabeled control is cells without incubation with primary and secondary antibodies (blue line).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD11a antibody [EP1285Y] (AB52895)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling CD11a with ab52895 at 1/100 dilution. Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/400 dilution was used as the secondary antibody (green). Confocal image shows membrane and cytoplasmic staining on Jurkat cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows;
1. ab52895 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11a antibody [EP1285Y] (AB52895)

Immunohistochemical analysis of paraffin-embedded Human squamous cell cervical carcinoma labeling CD11a with ab52895 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab79051) at 1/500 dilution. Membrane/cytoplasmic staining on stromal inflammatory cells of human cervical cancer is observed. The negative control utilised PBS instead of primary antibody. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-CD11a antibody [EP1285Y] (AB52895)

CD11a was immunoprecipitated from 1mg of Jurkat(Human T cell leukemia cells from peripheral blood) whole cell lysates using ab52895 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab52895 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1000 dilution.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-CD11a antibody [EP1285Y] (ab52895)

Predicted band size: 128 kDa

Observed band size: 180 kDa

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• WB

Supplier Data

Western blot - Anti-CD11a antibody [EP1285Y] (AB52895)

Blocking and Dilution buffer : 5% NFDM/TBST

The band in THP-1 cells is higher than that in Jurket cells due to the different glycosylation level in different materials.

All lanes:

Western blot - Anti-CD11a antibody [EP1285Y] (ab52895) at 1/5000 dilution

Lane 1:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg

Lane 2:

THP-1 (Human monocytic leukemia cells) whole cell lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 128 kDa

Observed band size: 180 kDa

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