CD11b/c Antibody, anti-rat, REAfinity™

Name: CD11b/c Antibody, anti-rat, REAfinity™
Availability: InStock

CD11b/c Antibody, anti-rat, REAfinity™Extended ValidationRecombinant

Clone:

REA325

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

ITGAM, ITGAX, Leukocyte adhesion glycoprotein p150,95 α chain (Leukocyte adhesion receptor p150,95), RGD1561123, CR-3 α chain, Cell surface glycoprotein MAC-1 subunit α, Leukocyte adhesion receptor MO1, Neutrophil adherence receptor, CD11b

Flow cytometry applications forCD11b/c Antibody, anti-rat, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 770
pure

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:

130-120-288, 130-120-214

Protocols:

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

Description:

Splenocytes from Lewis rats were stained with CD11b/c antibodies or with the corresponding REA Control (S) antibodies (left images) and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Extended validation for CD11b/c Antibody, anti-rat, 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 REA325
OX-42	++
WT.5 (only CD11b)	-
Cells were incubated with an excess of purified unconjugated CD11b/c (REA325) 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 CD11b/c. Splenocytes from Wistar rat were stained with CD11b/c antibodies and with a suitable counterstaining. As a control, CD11b/c 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 CD11b/c. Splenocytes from Wistar rat were stained with CD11b/c antibodies and with a suitable counterstaining. As a control, CD11b/c 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 CD11b/c. Splenocytes from Wistar rat were stained with CD11b/c antibodies and with a suitable counterstaining. As a control, CD11b/c 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 CD11b/c. Splenocytes from Wistar rat were stained with CD11b/c antibodies and with a suitable counterstaining. As a control, CD11b/c 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 CD11b/c (REA325). 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 CD11b/c (REA325). 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 CD11b/c Antibody, anti-rat, REAfinity™

Overview

Clone REA325 recognizes the rat CD11b and c antigen, which are both single-pass type I membrane glycoproteins also known as integrin α-M (CD11b) and integrin α-X (CD11c). CD11c and CD11b are expressed on monocytes, granulocytes, macrophages, dendritic cells, NK cells, and a subset of lymphocytes.

Additional information: Clone REA325 displays negligible binding to Fc receptors.

Alternative names

ITGAM, ITGAX, Leukocyte adhesion glycoprotein p150,95 α chain (Leukocyte adhesion receptor p150,95), RGD1561123, CR-3 α chain, Cell surface glycoprotein MAC-1 subunit α, Leukocyte adhesion receptor MO1, Neutrophil adherence receptor, CD11b

Detailed product information

Technical specifications

Clone	REA325
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	rat
Antigen	CD11b/c
Alternative names of antigen	ITGAM, ITGAX, Leukocyte adhesion glycoprotein p150,95 α chain (Leukocyte adhesion receptor p150,95), RGD1561123, CR-3 α chain, Cell surface glycoprotein MAC-1 subunit α, Leukocyte adhesion receptor MO1, Neutrophil adherence receptor, CD11b
Molecular mass of antigen [kDa]	128
Distribution of antigen	monocytes, macrophages, granulocytes, dendritic cells, NK cells, lymphocytes
Entrez Gene ID	25021, 499271
RRID	AB_2752041, AB_2752218, AB_2752215, AB_2811490, AB_2857720, AB_2904693, AB_2876964, AB_2654680, AB_2654681, AB_2654692, AB_2752052

Resources for CD11b/c Antibody, anti-rat, REAfinity™

Certificates

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

References for CD11b/c Antibody, anti-rat, REAfinity™

Publications

Hubert, F. X. et al. (2004)
Rat plasmacytoid dendritic cells are an abundant subset of MHC Class II
⁺
CD4
⁺
CD11b
^–
OX62
^–
and type I IFN-producing cells that exhibit selective expression of Toll-like receptors 7 and 9 and strong responsiveness to CpG.
J Immunol 172(12): 7485-7494
Kriegel, M. A. et al. (2012)
Pancreatic islet expression of chemokine CCL2 suppresses autoimmune diabetes via tolerogenic CD11c
⁺
CD11b
⁺
dendritic cells.
Proc. Natl. Acad. Sci. U.S.A. 109(9): 3457-3462
Milling, S. et al. (2010) Subsets of migrating intestinal dendritic cells. Immunol. Rev. 234(1): 259-267

Applications

Extended validation