CD11c Antibody, anti-mouse, REAfinity™

Name: CD11c Antibody, anti-mouse, REAfinity™
Availability: InStock

CD11c Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA754

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

p150, integrin αX, ITGAX, CR4

Flow cytometry applications forCD11c Antibody, anti-mouse, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 770
PerCP-Vio 700
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-110-839, 130-110-702

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Splenocytes from C57BL/6 mice were stained with CD11c antibodies or with the corresponding REA Control antibodies (left image) as well as with Anti-MHC Class II antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.

MICS (MACSima Imaging Cyclic Staining) applications forCD11c Antibody, anti-mouse, REAfinity™

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-110-839, 130-110-702

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-110-839, 130-110-702

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD11c Antibody, anti-mouse, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA754
N418	++
HL3	++
Cells were incubated with an excess of purified unconjugated CD11c (REA754) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for CD11c. Splenocytes from C57BL/6 mice were stained with CD11c antibodies and plotted against the side scatter. As a control, CD11c antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD11c. Splenocytes from C57BL/6 mice were stained with CD11c antibodies and plotted against the side scatter. As a control, CD11c antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD11c. Splenocytes from C57BL/6 mice were stained with CD11c antibodies and plotted against the side scatter. As a control, CD11c antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD11c. Splenocytes from C57BL/6 mice were stained with CD11c antibodies and plotted against the side scatter. As a control, CD11c antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD11c. Splenocytes from C57BL/6 mice were stained with CD11c antibodies and plotted against the side scatter. As a control, CD11c antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD11c (REA754). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD11c (REA754). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD11c Antibody, anti-mouse, REAfinity™

Overview

Clone REA754 recognizes the mouse CD11c antigen, also known as integrin αX or Itgax. CD11c is present in dendritic cells in lymphoid organs and blood, in Langerhans cells in the epidermis, in dendritic cell progenitors in the bone marrow, and in

in vitro

–generated bone marrow–derived dendritic cells. In spleen and lymph node, CD11c is found at high levels on conventional CD11c

⁺

CD45R

^–

mPDCA-1

^–

dendritic cells, and at moderate levels on CD11c

⁺

CD45R

⁺

mPDCA-1

⁺

plasmacytoid dendritic cells. CD11c is reported to be weakly expressed on NK cells, B cells, and T cell subsets. About 1–3% of splenocytes, 2% of bone marrow cells, as well as <1% of lymph node cells and thymocytes express CD11c. CD11c is a receptor for fibrinogen. It mediates cell-cell interaction during inflammatory responses and is involved in monocyte adhesion and chemotaxis.

Additional information: Clone REA754 displays negligible binding to Fc receptors.

Alternative names

p150, integrin αX, ITGAX, CR4

Detailed product information

Technical specifications

Clone	REA754
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody, human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	mouse
Antigen	CD11c
Alternative names of antigen	p150, integrin αX, ITGAX, CR4
Molecular mass of antigen [kDa]	127
Distribution of antigen	T cells, NK cells, B cells, dendritic cells, granulocytes, macrophages, monocytes, lymphocytes
Entrez Gene ID	16411
RRID	AB_2654706, AB_2654707, AB_2654708, AB_2654709, AB_2654710, AB_2654711, AB_2654712, AB_2654713, AB_2654714, AB_2654715, AB_2654716, AB_2654717, AB_2654718, AB_2654719, AB_2654705

Resources for CD11c Antibody, anti-mouse, REAfinity™

Documents and Protocols

App notes/customer reports

REAfinity™ Recombinant Antibodies - Background-free analysis of mouse TILs

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD11c Antibody, anti-mouse, REAfinity™

Publications

Springer, T. A. (1994) Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell 76(2): 301-314
López-Rodríguez, C. et al. (1996) AP-1 regulates the basal and developmentally induced transcription of the CD11c leukocyte integrin gene. J Immunol 156(10): 3780-3787

Applications

Extended validation