Clone:
REA422
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
LY6A, Ly-6A.2, Ly-6A/E, Ly-6E.1, TAP

Extended validation for Sca-1 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 REA422
D7++
E13-161.7++
Cells were incubated with an excess of purified unconjugated Sca-1 (REA422) 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 Sca-1. Bone marrow cells from C57BL/6 mice were stained with Sca-1 antibodies and with a suitable counterstaining. As a control, Sca-1 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 Sca-1. Bone marrow cells from C57BL/6 mice were stained with Sca-1 antibodies and with a suitable counterstaining. As a control, Sca-1 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 Sca-1. Bone marrow cells from C57BL/6 mice were stained with Sca-1 antibodies and with a suitable counterstaining. As a control, Sca-1 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 Sca-1. Bone marrow cells from C57BL/6 mice were stained with Sca-1 antibodies and with a suitable counterstaining. As a control, Sca-1 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 Sca-1. Bone marrow cells from C57BL/6 mice were stained with Sca-1 antibodies and with a suitable counterstaining. As a control, Sca-1 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 Sca-1 (REA422). 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 Sca-1 (REA422). 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 Sca-1 (REA422). 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 Sca-1 Antibody, anti-mouse, REAfinity™

Overview

Clone REA422 recognizes the mouse stem cell antigen-1 (SCA-1) antigen, a 18 kDa glycosylphosphatidylinositol (GPI) linked protein which is also known as lymphocyte antigen 6A and 6E (Ly-6A/E). Clone REA422 specifically recognizes both Ly-6E.1 and Ly-6A.2, which are gene products of the two Ly-6A/E alleles expressed in different mouse strains (Ly-6E.1: e.g. BALB/c, C3H, NZB; Ly-6A.2: e.g. C57BL/6, SJL, 129, AKR). Sca-1 is expressed on hematopoietic stem cells and progenitor cells in mouse bone marrow and is one of the defining markers for so-called KSL (c-kit
+
Sca-1
+
Lin
-
) cells. Sca-1 in combination with CD105 (Endoglin) expression has been used to define long-term repopulating hematopoietic stem cells (LTR-HSCs) in mouse bone marrow. In addition, Sca-1
+
bone marrow cells also contain the mesenchymal stem cell fraction. Sca-1
+
cells have been shown to give rise to hepatocytes
in vivo
and neural cells
in vitro
. Furthermore, Sca-1 is expressed on stem cells in a variety of nonhematopoietic tissues, such as adult liver, heart, and prostate.
Additional information: Clone REA422 displays negligible binding to Fc receptors.

Alternative names

LY6A, Ly-6A.2, Ly-6A/E, Ly-6E.1, TAP

Detailed product information

Technical specifications

CloneREA422
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenSca-1
Alternative names of antigenLY6A, Ly-6A.2, Ly-6A/E, Ly-6E.1, TAP
Molecular mass of antigen [kDa]9
Distribution of antigenstem cells, hematopoietic stem and progenitor cells
Entrez Gene ID110454
RRIDAB_2727567, AB_2727592, AB_2811562, AB_2905382, AB_2889749, AB_2653396, AB_2921945, AB_2751322

Resources for Sca-1 Antibody, anti-mouse, REAfinity™

Documents and Protocols

Certificates

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

References for Sca-1 Antibody, anti-mouse, REAfinity™

Publications

  1. Bradfute, S. B. et al. (2005) Roles of Sca-1 in hematopoietic stem/progenitor cell function. Exp. Hematol. 33(7): 836-843
  2. Khan, K. D. et al. (1990) Characterization of promoter elements of an interferon-inducible Ly-6E/A differentiation antigen, which is expressed on activated T cells and hematopoietic stem cells. Mol. Cell. Biol. 10(10): 5150-5159
  3. Houlihan, D. D. et al. (2012) Isolation of mouse mesenchymal stem cells on the basis of expression of Sca-1 and PDGFR-α. Nat. Protoc. 7(12): 2103-2111

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