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

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 clonesOverlap in epitope recognition with 14.8
REA639+
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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
View details
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.
View details
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

Clone14.8
Clonalitymonoclonal
Isotyperat IgG2bκ
Isotype controlIsotype Control Antibody, rat IgG2b
Hostrat
Type of antibodyPrimary antibodies
Speciesmouse
AntigenCD45RA
Alternative names of antigenPtprc, loc, Lyt-4, CD45, L-CA, Ly-5
Molecular mass of antigen [kDa]135(calculated based on gene prediction)
Distribution of antigenB cells, NK cells, monocytes, T cells, thymocytes
Entrez Gene ID19264
RRIDAB_2658339, AB_2658340, AB_2658341, AB_2658342, 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

  1. 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
  2. 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
  3. Trowbridge, I. S. et al. (1991) CD45: a leukocyte-specific member of the protein tyrosine phosphatase family. J. Immunol. 1095(1): 46-56

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