CD45RA Antibody, anti-mouse

Name: CD45RA Antibody, anti-mouse
Availability: InStock

CD45RA Antibody, anti-mouseExtended Validation

Clone:

14.8

Type of antibody:

Primary antibodies

Isotype:

rat IgG2bκ

Applications:

FC, MICS, IF, IHC

Alternative names:

Ptprc, loc, Lyt-4, CD45, L-CA, Ly-5

Flow cytometry applications forCD45RA Antibody, anti-mouse

APC
Biotin
FITC
PE
Vio Bright B515

Conjugate:

APC

Recommended dilution:

1:10

Remark:

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

Tested:

QC tested

Products used:

130-102-484

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:10)

Description:

Spleen cells of BALB/c mice were stained with CD45RA antibodies, as well as with CD19 antibodies and analyzed by flow cytometry using the MACSQuant^® Analyzer. Cells labeled with Biotin were stained with Anti-Biotin-PE in addition. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD45RA Antibody, anti-mouse

APC
FITC
PE

Conjugate:

APC

Recommended dilution:

1:10

Fixation:

Acetone

Tested:

during development

Species tested:

mouse

Products used:

130-102-484

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Extended validation for CD45RA Antibody, anti-mouse

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 14.8
REA639	+ (PE)
REA639	++ (FITC)
Cells were incubated with an excess of purified unconjugated CD45RA (14.8) 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA. C57BL/6 Mouse splenocytes were stained with CD45RA antibodies and with a suitable counterstaining. As a control, CD45RA 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 CD45RA (14.8). 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 CD45RA (14.8). 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 CD45RA Antibody, anti-mouse

Overview

The monoclonal antibody 14.8 recognizes the exon A-dependent epitope CD45RA, one of eight isoforms of the transmembrane glycoprotein CD45, which is a member of the PTP (Protein Tyrosine Phosphatase) family. CD45RA is expressed highly on B cells, weakly on CD8

⁺

T cells and nearly on all B-lineage cells but it is not detectable on hematopoietic stem cells or myeloid progenitors.

Alternative names

Ptprc, loc, Lyt-4, CD45, L-CA, Ly-5

Detailed product information

Technical specifications

Clone	14.8
Clonality	monoclonal
Isotype	rat IgG2bκ
Isotype control	Isotype Control Antibody, rat IgG2b
Host	rat
Type of antibody	Primary antibodies
Species	mouse
Antigen	CD45RA
Alternative names of antigen	Ptprc, loc, Lyt-4, CD45, L-CA, Ly-5
Molecular mass of antigen [kDa]	135(calculated based on gene prediction)
Distribution of antigen	B cells, NK cells, monocytes, T cells, thymocytes
Entrez Gene ID	19264
RRID	AB_2658339, AB_2658340, AB_2658341, AB_2658342, AB_2922065, AB_2658338

Resources for CD45RA Antibody, anti-mouse

Certificates

Please follow this

link

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

References for CD45RA Antibody, anti-mouse

Publications

Hathcock, K. S. et al. (1992) Expression of variable exon A-, B-, and C-specific CD45 determinants on peripheral and thymic T cell populations. J Immunol 148: 19-28
Inoue, T. et al. (1993)
Distinction of mouse CD8
⁺
suppressor effector T cell clones from cytotoxic T cell clones by cytokine.
J Immunol 150: 2121-2128
Trowbridge, I. S. et al. (1991) CD45: a leukocyte-specific member of the protein tyrosine phosphatase family. J Immunol 1095(1): 46-56

Applications

Extended validation