Clone:
REA364
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IHC, IF
Alternative names:
Spn, B-cell differentiation antigen LP-3, GALGP, Ly-48, Sialophorin, CD43

Extended validation for CD43 Glycoform 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 clonesOverlap in epitope recognition with REA364
eBioR2/60-
1B11++
S11-
S7-
Cells were incubated with an excess of purified unconjugated CD43 Glycoform (REA364) 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.
View details
Flow cytometric comparison of different clones for ( CD43 Glycoform specificity from PIM). C57BL/6 Mouse splenocytes were stained with CD43 Glycoform antibodies and with a suitable counterstaining. As a control, CD43 Glycoform 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.
View details
Flow cytometric comparison of different clones for ( CD43 Glycoform specificity from PIM). C57BL/6 Mouse splenocytes were stained with CD43 Glycoform antibodies and with a suitable counterstaining. As a control, CD43 Glycoform 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.
View details
Flow cytometric comparison of different clones for ( CD43 Glycoform specificity from PIM). C57BL/6 Mouse splenocytes were stained with CD43 Glycoform antibodies and with a suitable counterstaining. As a control, CD43 Glycoform 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 Glycoform specificity from PIM). C57BL/6 Mouse splenocytes were stained with CD43 Glycoform antibodies and with a suitable counterstaining. As a control, CD43 Glycoform 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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD43 Glycoform (REA364). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD43 Glycoform (REA364). 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 Glycoform (REA364). 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 Glycoform Antibody, anti-mouse, REAfinity™

Overview

Clone REA364 recognizes the activation-associated glycoform (130-kD glycoform) of the murine CD43 antigen, a cell-surface glycoprotein which is also known as sialophorin or leukosialin. CD43 is a heavily O-glycosylated protein which is expressed in a variety of hematopoietic lineage cells, including T lymphocytes. It has been reported to have two glycoforms, one with a molecular mass of 115 kD that is expressed on all T cells and one with a mass of 130 kD that is expressed only on activated T cells. Biochemical analysis showed that CD43 on resting T cells carries tetrasaccharides (115-kD glycoform), whereas activated T cells carry branched hexasaccharide core two O-glycans (130-kD glycoform). Expression of the 130-kD glycoform of CD43 on all T cells in transgenic mice shows a reduction in primary T-cell immune responses that is probably due to the antiadhesive function of this glycoform.
Additional information: Clone REA364 displays negligible binding to Fc receptors.

Alternative names

Spn, B-cell differentiation antigen LP-3, GALGP, Ly-48, Sialophorin, CD43

Detailed product information

Technical specifications

CloneREA364
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD43 Glycoform
Alternative names of antigenSpn, B-cell differentiation antigen LP-3, GALGP, Ly-48, Sialophorin, CD43
Molecular mass of antigen [kDa]38
Distribution of antigenB cells, T cells, lymphocytes
Entrez Gene ID20737
RRIDAB_2658142, AB_2658143, AB_2658144, AB_2658145, AB_2658148, AB_2658149, AB_2905011

Resources for CD43 Glycoform Antibody, anti-mouse, REAfinity™

Certificates

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References for CD43 Glycoform Antibody, anti-mouse, REAfinity™

Publications

  1. Carlow, D. A. et al. (1999) A novel CD8 T cell-restricted CD45RB epitope shared by CD43 is differentially affected by glycosylation J. Immunol. 163(3): 1441-1448
  2. 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: 3637- 3640
  3. Alcaide, P. et al. (2007)
    The 130-kDa glycoform of CD43 functions as an E-selectin ligand for activated Tʜ1 cells
    in vitro
    and in delayed-type hypersensitivity reactions
    in vivo.
    J. Invest. Dermatol. 127(8): 1964-1972

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