Recombinant Anti-CD11c antibody [EP1347Y] - C-terminal (ab52632)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD11c with Purified ab52632 at 1:500 dilution (0.21 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

Blocking and diluting buffer: 5% NFDM/TBST

ab52632 (purified) at 1:20 dilution (2µg) immunoprecipitating CD11c in THP-1 (Human monocytic leukemia monocyte) starved for 2 hours, then treated with Phorbol-12-myristate-13-acetate at 100ng/ml for 96 hours whole cell lysate.

Lane 1 (input): THP-1 (Human monocytic leukemia monocyte) starved for 2 hours, then treated with Phorbol-12-myristate-13-acetate at 100ng/ml for 96 hours whole cell lysate 10ug
Lane 2 (+): ab52632 & THP-1 (Human monocytic leukemia monocyte) starved for 2 hours, then treated with Phorbol-12-myristate-13-acetate at 100ng/ml for 96 hours whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52632 in THP-1 (Human monocytic leukemia monocyte) starved for 2 hours, then treated with Phorbol-12-myristate-13-acetate at 100ng/ml for 96 hours whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST."

Immunohsitochemical analysis of paraffin-embedded human tonsil tissue labelling CD11c with ab52632 at 1/500 dilution. Tissue fixed with 10% NBF. An Alexa Fluor® 647 Goat anti-Rabbit IgG secondary antibody was used. Heat mediated antigen retrieval with Tris/EDTA, ph 9 carried out.

Unpurified ab52632 showing positive staining in Normal tonsil tissue.

Unpurified ab52632 at 1/400 dilution staining CD11c in human tonsil tissue by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Sections were paraformaldehyde fixed, prior to blocking in 10% BSA in FCS (20ml) + DMEM (80ml) for 30 minutes at 21°C and then incubated with ab52632 for 20 hours at 4°C. A biotin conjugated rabbit polyclonal to mouse Ig, diluted 1/400, was used as the secondary antibody.

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Unpurified ab52632 showing negative staining in Normal brain tissue.

This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab52632 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

Unpurified ab52632 showing negative staining in Normal breast tissue.

Unpurified ab52632 showing negative staining in Skeletal muscle tissue.

Unpurified ab52632 showing positive staining in Normal spleen tissue.