Anti-CD11c antibody [EP1347Y] - C-terminal

• IHC-P

AbReview69857****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

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.

Image courtesy of an anonymous abreview

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Tissue Microarrays stained for "Anti-CD11c antibody [EP1347Y] - C-terminal" using "ab52632"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab52632 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Unpurified ab52632 showing negative staining in Normal breast tissue.

• IHC

Lab

Immunohistochemistry - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling CD11c with ab52632 at a concentration of 0.08 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32mins.

ab52632 anti-CD11c [EP1347Y] was incubated for 16 mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

AbReview16265****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

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.

This image is courtesy of an Abreview submitted by James Jupp

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human Hodgkin's lymphoma sections labeling CD11c with ab52632 at 1 : 5000 dilution (0.02 μg/ml). Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) (ready to use) secondary antibody was used. Positive staining on human Hodgkin's lymphoma. The section was incubated with ab52632 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human spleen tissue sections labeling CD11c with ab52632 at 1 : 5000 dilution (0.02 μg/ml). Heat mediated antigen retrieval was performed using Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) (ready to use) secondary antibody was used. Positive staining on human spleen. The section was incubated with ab52632 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IP

Unknown

Immunoprecipitation - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

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."

All lanes:

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

Predicted band size: 127 kDa

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• WB

Lab

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (ab52632) at 1/1000 dilution

Lane 1:

Untreated THP-1 (Human monocytic leukemia monocyte) whole cell lysates at 15 µg

Lane 2:

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 lysates at 15 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: 127 kDa

false

• WB

Lab

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

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.

All lanes:

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (ab52632) at 1/1000 dilution

Lane 1:

THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate at 10 µg

Lane 2:

THP1 (Human acute monocytic leukemia cell line) + PMA 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 1/10000 dilution

Predicted band size: 127 kDa

Observed band size: 150 kDa

true

Exposure time: 4min

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

Unpurified ab52632 showing negative staining in Skeletal muscle tissue.

• WB

Unknown

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

All lanes:

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (ab52632) at 1/1000 dilution

All lanes:

THP-1 + TPA lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 127 kDa

Observed band size: 150 kDa

false

• WB

CiteAb

Western blot - Anti-CD11c antibody [EP1347Y] - C-terminal (AB52632)

CD11c western blot using anti-CD11c antibody [EP1347Y] - C-terminal ab52632. Publication image and figure legend from Francistiová, L., Vörös, K., et al., 2022, Front Mol Neurosci, PubMed 35095416.

ab52632 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab52632 please see the product overview.

Representative confocal images suggest intracellular localization of the P2X7 receptor’s signal (in green) in Ctrl and fAD neuronal cells (NPC or neurons; labeled with TUBB3 (in red) (A) and membranous localization in microglia-like cells (B). Each staining is a representative picture of at least six independent experiments. Microglia-like cells were co-stained with IBA1 (red). Scale bar : 20 μm (C,D) and (E) show representative Western blot results of biotinylation analysis. Arrows indicate the bands representing the canonical ≈72 kDa sized P2X7R localized in the IC (intra-cellular) fraction in the case of Ctrl and fAD neurons’ (TD53) samples (D) and the M (membrane) fraction of microglia samples (E) and both fractions in NPC samples (C). Red arrows indicate the presence of the P2X7R band in the M fractions. The Integrin α7 (C,D) and CD11c (E) are plasma membrane proteins present only in the M factions. HSP27 is a nuclear protein present only in the IC fractions (C–E). Integrin α7 and HSP27 were used as controls of the efficiency of biotinylation and the separation of biotinylated proteins. Western blot measurements were per-formed as biological triplicates and a duplicate in the case of microglia-like cells. Ctrl, control; Neu, neuron; NPC, neuronal progenitor cell; IC, intra-cellular; M, membrane.

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