CD45RC Antibody, anti-rat, REAfinity™

Name: CD45RC Antibody, anti-rat, REAfinity™
Availability: InStock

CD45RC Antibody, anti-rat, REAfinity™Extended ValidationRecombinant

Clone:

REA1000

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC

Alternative names:

CD45, LCA, L-CA

Flow cytometry applications forCD45RC Antibody, anti-rat, REAfinity™

APC
APC-Vio 770
Biotin
FITC
PE
PE-Vio 615
PE-Vio 770
Vio Bright B515
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:

130-116-900, 130-116-780

Protocols:

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

Description:

Splenocytes from Sprague Dawley^® rats were stained with CD45RC antibodies or with the corresponding REA Control antibodies (left images) as well as with CD3 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.

Extended validation for CD45RC 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 REA1000
OX-22	++
Cells were incubated with an excess of purified unconjugated CD45RC (REA1000) 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC. Wistar rat Splenocytes were stained with CD45RC antibodies and with a suitable counterstaining. As a control, CD45RC 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 CD45RC (REA1000). 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 CD45RC (REA1000). 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 CD45RC (REA1000). 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 CD45RC (REA1000). 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 CD45RC Antibody, anti-rat, REAfinity™

Overview

Clone REA1000 recognizes the RC isoform of the rat CD45 antigen, also known as leukocyte common antigen (LCA), which is a single-chain type I transmembrane glycoprotein. CD45 has multiple isoforms and differential glycosylation, CD45RC is the high molecular weight form of CD45. CD45 is expressed at high levels on all cells of hematopoietic origin except for erythrocyte and platelets. It plays a role in signal transduction through B and T cell antigen receptors. The CD45RC isoform is expressed on B cells, most cytotoxic T cells, and a subset of T helper cells, but not expressed on myeloid cells.

Additional information: Clone REA1000 displays negligible binding to Fc receptors.

Alternative names

CD45, LCA, L-CA

Detailed product information

Technical specifications

Clone	REA1000
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	CD45RC
Alternative names of antigen	CD45, LCA, L-CA
Molecular mass of antigen [kDa]	141
Distribution of antigen	B cells, T cells, CD4+ T cells, CD8+ T cells
Entrez Gene ID	24699
RRID	AB_2727752, AB_2727683, AB_2727753, AB_2727684, AB_2727754, AB_2727755, AB_2727685, AB_2727756, AB_2727757, AB_2727758, AB_2727686, AB_2727759, AB_2727751

Resources for CD45RC Antibody, anti-rat, REAfinity™

Certificates

Please follow this

link

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

References for CD45RC Antibody, anti-rat, REAfinity™

Publications

Sunderland, C. A. et al. (1979) Purification with monoclonal antibody of a predominant leukocyte-common antigen and glycoprotein from rat thymocytes. Eur. J. Immunol. 9(2): 155-159
Mojcik, D. L. et al. (1987) Monoclonal antibodies to RT7 and LCA antigens in the rat: cell distribution and segregation analysis. Hybridoma 6(5): 531-543

Applications

Extended validation