Clone:
REA421
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
Ly-1, Ly-12, Ly-A, Lyt-1

Extended validation for CD5 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 REA421
53-7.3++
Cells were incubated with an excess of purified unconjugated CD5 (REA421) 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 CD5. Splenocytes from CD1 mice were stained with CD5 antibodies and with a suitable counterstaining. As a control, CD5 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 CD5. Splenocytes from CD1 mice were stained with CD5 antibodies and with a suitable counterstaining. As a control, CD5 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 CD5. Splenocytes from CD1 mice were stained with CD5 antibodies and with a suitable counterstaining. As a control, CD5 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 CD5. Splenocytes from CD1 mice were stained with CD5 antibodies and with a suitable counterstaining. As a control, CD5 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 CD5. Splenocytes from CD1 mice were stained with CD5 antibodies and with a suitable counterstaining. As a control, CD5 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 CD5 (REA421). 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 CD5 (REA421). 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 CD5 (REA421). 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 CD5 Antibody, anti-mouse, REAfinity™

Overview

Clone REA421 recognizes the mouse CD5 antigen, a 67 kDa single-pass type I membrane protein which is also known as lymphocyte antigen 1 (Ly-1). CD5 is expressed by a majority of thymocytes, mature T cells, and a subset of B cells. The expression of CD5 by a small B cell subset characterizes a developmentally and functionally distinct lineage of B cells called B-1a cells. CD5 plays a role in the interaction of T and B cells and is a counter-receptor for CD72.
Additional information: Clone REA421 displays negligible binding to Fc receptors.

Alternative names

Ly-1, Ly-12, Ly-A, Lyt-1

Detailed product information

Technical specifications

CloneREA421
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD5
Alternative names of antigenLy-1, Ly-12, Ly-A, Lyt-1
Molecular mass of antigen [kDa]52
Distribution of antigenB cells, T cells
Entrez Gene ID12507
RRIDAB_2752044, AB_2752055, AB_2752026, AB_2811489, AB_2658566, AB_2658567, AB_2658578, AB_2658579, AB_2658580, AB_2658581, AB_2752057

Resources for CD5 Antibody, anti-mouse, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for CD5 Antibody, anti-mouse, REAfinity™

Publications

  1. Tarakhovsky, A. et al. (1995) A role for CD5 in TCR-mediated signal transduction and thymocyte selection. Science 269(5223): 535-537
  2. Efanov, A. et al. (2010)
    CD5
    +
    CD23
    +
    leukemic cell populations in TCL1 transgenic mice show significantly increased proliferation and Akt phosphorylation.
    Leukemia 24(5): 970-975
  3. Lundy, S. K. et al. (2014)
    Characterization and activity of Fas ligand producing CD5
    +
    B cells.
    Methods Mol. Biol. 1190: 81-102

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