CD43 Antibody, anti-mouse, REAfinity™

Name: CD43 Antibody, anti-mouse, REAfinity™
Availability: InStock

CD43 Antibody, anti-mouse, REAfinity™Extended ValidationRecombinant

Clone:

REA840

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC

Alternative names:

Spn, Leukosialin, GALGP, Ly-48, gpL115, Sialophorin

Flow cytometry applications forCD43 Antibody, anti-mouse, REAfinity™

APC
APC-Vio 770
PE
PE-Vio 615
PE-Vio 770
PerCP-Vio 700
VioBlue

Conjugate:

APC

Recommended dilution:

1:50

Remark:

The antibody is suited for staining of formaldehyde-fixed cells.

Tested:

QC tested

Products used:

130-112-888, 130-112-699

Protocols:

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

Description:

Splenocytes of C57BL/6 mice were stained with CD43 antibodies or with the corresponding REA Control antibodies (left image) as well as with CD19 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 forCD43 Antibody, anti-mouse, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

mouse

Products used:

130-112-888, 130-112-699

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:

130-112-888, 130-112-699

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD43 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 REA840
S7	++
S11	-
1B11	-
L11	++
Cells were incubated with an excess of purified unconjugated CD43 (REA840) 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 CD43. Splenocytes from C57BL/6 were stained with CD43 antibodies and with a suitable counterstaining. As a control, CD43 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 CD43. Splenocytes from C57BL/6 were stained with CD43 antibodies and with a suitable counterstaining. As a control, CD43 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 CD43. Splenocytes from C57BL/6 were stained with CD43 antibodies and with a suitable counterstaining. As a control, CD43 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 CD43. Splenocytes from C57BL/6 were stained with CD43 antibodies and with a suitable counterstaining. As a control, CD43 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 CD43. Splenocytes from C57BL/6 were stained with CD43 antibodies and with a suitable counterstaining. As a control, CD43 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 CD43 (REA840). 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 CD43 (REA840). 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 CD43 Antibody, anti-mouse, REAfinity™

Overview

Clone REA840 recognizes the mouse CD43 antigen, a glycosylated protein also known as leukosialin or sialophorin, which is expressed on IL-7–responsive pro–B cells, plasma cells, peritoneal and spleen CD5

⁺

B cells (B-1 cells), granulocytes, monocytes, macrophages, platelets, NK cells, thymocytes, peripheral cytotoxic T cells, and most T helper cells. CD43 is not expressed on resting conventional peripheral B cells. Thus, resting B cells are easily identified by the lack of CD43 expression.

Additional information: Clone REA840 displays negligible binding to Fc receptors.

Alternative names

Spn, Leukosialin, GALGP, Ly-48, gpL115, Sialophorin

Detailed product information

Technical specifications

Clone	REA840
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	CD43
Alternative names of antigen	Spn, Leukosialin, GALGP, Ly-48, gpL115, Sialophorin
Molecular mass of antigen [kDa]	38
Distribution of antigen	B cells, granulocytes, lymphocytes, macrophages, megakaryocytes, stem cells, monocytes, NK cells, platelets, red blood cells, T cells, basophils, mesenchymal stem cells, neutrophils, plasma cells, thymocytes, bone marrow
Entrez Gene ID	20737
RRID	AB_2658115, AB_2658117, AB_2658118, AB_2658119, AB_2658120, AB_2658121, AB_2658123, AB_2658124, AB_2905010, AB_2658114, AB_2658116, AB_2658122, AB_2658113

Resources for CD43 Antibody, anti-mouse, REAfinity™

Certificates

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

References for CD43 Antibody, anti-mouse, REAfinity™

Publications

Cyster, J. et al. (1990) Protein sequence and gene structure for mouse leukosialin (CD43), a T lymphocyte mucin without introns in the coding sequence. Eur. J. Immunol. 20(4): 875-881
Dorfman, K. S. et al. (1990) The nucleotide sequence of Ly 48 (mouse leukosialin, sialophorin): the mouse homolog of CD43. Nucleic Acids Res. 18(16): 4932-4932
van den Berg, T. K. et al. (2001) Cutting Edge: CD43 functions as a T cell counterreceptor for the macrophage adhesion receptor Sialoadhesin (Siglec-1). J Immunol 166(6): 3637-3640

Applications

Extended validation