Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Mouse, Rat, Transgenic Mouse

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Sca-1/Ly6 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Sca-1 C-terminally truncated Ly-6E allele
Leu27-Gly119
Accession # CAA28351

Specificity

Detects mouse Sca-1/Ly6 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse Sca‑1/Ly6 Antibody

Sca‑1/Ly6 antibody in Mouse Splenocytes by Immunocytochemistry (ICC).

Sca‑1/Ly6 in Mouse Splenocytes.

Sca-1/Ly6 was detected in immersion fixed mouse splenocytes using 10 µg/mL Goat Anti-Mouse Sca-1/Ly6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1226) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Mouse Sca-1/Ly6 by Western Blot

Detection of Mouse Sca-1/Ly6 by Western Blot

Sca1, a marker of mammary stem/progenitor cells, is increased in DNIIR and Wnt5a-/- glands. mRNA and protein were extracted from several wild type (WT) and MT-DNIIR (DNIIR) mice. (a) Sca1 mRNA levels were determined by semi-quantitative RT-PCR and (b) protein levels were determined by western blot. Sca1 levels were increased in DNIIR glands relative to controls. (c) Sca1 protein levels were determined in WT and Wnt5a null (-/-) glands using western blot analysis. Sca1 levels were increased in Wnt5a-null tissue relative to controls. 18S and beta -Tubulin ( beta -Tub) were used as normalisation controls. DNIIR = dominant-negative transforming growth factor-beta type II receptor; RT-PCR = reverse transcripase polymerase chain reaction; Wnt = Wingless-related protein family. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sca-1/Ly6 by Western Blot

Detection of Mouse Sca-1/Ly6 by Western Blot

Sca1, a marker of mammary stem/progenitor cells, is increased in DNIIR and Wnt5a-/- glands. mRNA and protein were extracted from several wild type (WT) and MT-DNIIR (DNIIR) mice. (a) Sca1 mRNA levels were determined by semi-quantitative RT-PCR and (b) protein levels were determined by western blot. Sca1 levels were increased in DNIIR glands relative to controls. (c) Sca1 protein levels were determined in WT and Wnt5a null (-/-) glands using western blot analysis. Sca1 levels were increased in Wnt5a-null tissue relative to controls. 18S and beta -Tubulin ( beta -Tub) were used as normalisation controls. DNIIR = dominant-negative transforming growth factor-beta type II receptor; RT-PCR = reverse transcripase polymerase chain reaction; Wnt = Wingless-related protein family. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sca-1/Ly6 by Western Blot

Detection of Mouse Sca-1/Ly6 by Western Blot

Ectopic expression of Wnt5a results in low expression of K6 and K14.Expression of molecular markers for basal and luminal progenitors in MMTV-Wnt1 and MMTV-Wnt1;MMTV-Wnt5a mammary glands was compared by western blot. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. Each lane contains protein isolated from a separate mouse. Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0113247), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sca-1/Ly6 by Western Blot

Detection of Mouse Sca-1/Ly6 by Western Blot

Sca1, a marker of mammary stem/progenitor cells, is increased in DNIIR and Wnt5a-/- glands. mRNA and protein were extracted from several wild type (WT) and MT-DNIIR (DNIIR) mice. (a) Sca1 mRNA levels were determined by semi-quantitative RT-PCR and (b) protein levels were determined by western blot. Sca1 levels were increased in DNIIR glands relative to controls. (c) Sca1 protein levels were determined in WT and Wnt5a null (-/-) glands using western blot analysis. Sca1 levels were increased in Wnt5a-null tissue relative to controls. 18S and beta -Tubulin ( beta -Tub) were used as normalisation controls. DNIIR = dominant-negative transforming growth factor-beta type II receptor; RT-PCR = reverse transcripase polymerase chain reaction; Wnt = Wingless-related protein family. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sca-1/Ly6 by Western Blot

Detection of Mouse Sca-1/Ly6 by Western Blot

Wnt5a expressing tumors demonstrate a decrease in markers of the basal tumor subtype.(A) Western blot using protein lysates isolated from the epithelium of MMTV-Wnt1 and MMTV-1;MMTV-Wnt5a mammary tumors. The expression of molecular markers of basal and luminal tumor subtypes were compared. Keratin 6 and Keratin 5 were strongly down-regulated in Wnt5a expressing tumors. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. (B–D) Immunostaining for K6. Sections from MMTV-Wnt1 (B) and MMTV-1;MMTV-Wnt5a (C) tumors were stained with anti-Keratin 6 antibody using immunofluorescence (K6 = green; nuclei = blue). The percentage of cells expressing K6 was determined and graphed (D). Values are means +/− standard error (n = 6 MMTV-Wnt1, 3 fields per tumor; n = 5 MMTV-Wnt1;MMTV-Wnt5a, 3 fields per tumor). MMTV-Wnt1;MMTV-Wnt5a tumors demonstrated a significant decrease in K6-expressing cells as measured by T-test (* = p<0.05). Image collected and cropped by CiteAb from the following open publication (https://dx.plos.org/10.1371/journal.pone.0113247), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sca-1/Ly6 by Western Blot

Detection of Mouse Sca-1/Ly6 by Western Blot

Sca1, a marker of mammary stem/progenitor cells, is increased in DNIIR and Wnt5a-/- glands. mRNA and protein were extracted from several wild type (WT) and MT-DNIIR (DNIIR) mice. (a) Sca1 mRNA levels were determined by semi-quantitative RT-PCR and (b) protein levels were determined by western blot. Sca1 levels were increased in DNIIR glands relative to controls. (c) Sca1 protein levels were determined in WT and Wnt5a null (-/-) glands using western blot analysis. Sca1 levels were increased in Wnt5a-null tissue relative to controls. 18S and beta -Tubulin ( beta -Tub) were used as normalisation controls. DNIIR = dominant-negative transforming growth factor-beta type II receptor; RT-PCR = reverse transcripase polymerase chain reaction; Wnt = Wingless-related protein family. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Sca-1/Ly6 by Immunohistochemistry

Detection of Mouse Sca-1/Ly6 by Immunohistochemistry

Sali spheres contain stem cells.(A) Sca-1 is present in the mouse submandibular gland on endothelial cells as well as on excretory and striated duct cells (D), and was clearly present at the onset of cultivation (D0) whereas acinar cells (AC) did not express Sca-1. At day 3 and later time-points, nearly all cells at the periphery of the Sali spheres showed high Sca-1 expression which decreased in time (D10). (B) Approximately 52.0+/−3.1% (Mean+/−STDEV) of cells in D3 cultured spheres expressed Sca-1, as quantified by flow cytometry. (C) c-Kit is only expressed by excretory duct cells (Tissue). Sali spheres showed similar c-Kit staining patterns as for Sca-1. (D) Most nuclei in excretory duct compartments and few nuclei in striated duct cells showed Musashi-1 presence (Tissue). Whereas Musashi-1 was still present in the nuclei of some cells at the onset of culturing (D0-arrow), most cells showed cytoplasmic localization which diminished in time (D3–5–10). Cells were visualized with DAPI (blue). Scale bar = 50 µm, inset = 20 µm. D = duct cells, AC = acinar cell, D0–3–5–10 represent days in culture. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/18446241), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse Sca‑1/Ly6 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Mouse splenocytes

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse splenocytes

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse Sca‑1/Ly6

Reviewed Applications

Read 1 review rated 5 using AF1226 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Sca-1/Ly6

This antibody reacts with Sca-1, an 18 kDa phosphatidylinositol-anchored protein that is a member of the lymphocyte antigen 6 (Ly-6) family (1). Sca-1 is encoded by the strain-specific Ly-6 E/A allelic gene. Its expression on multipotent hematopoietic stem cells (HSC) has been used as a marker of HSC in mice of both Ly‑6 haplotypes (2, 3). This antibody is frequently used in combination with lineage depletion antibodies to identify and isolate murine HSC. Sca-1-positive HSC can be found in the adult bone marrow, fetal liver and mobilized peripheral blood and spleen in the adult animal (2‑7). However, Sca-1 has also been discovered in several non‑hematopoietic tissues (1) and can be used to enrich progenitor cell populations other than HSC (8). It is suggested that Sca-1 could be involved in regulating both B and T cell activation (9‑12).

References

  1. Van de Rijn, M. et al. (1989) Proc. Natl.Acad. Sci. USA 86:4634.
  2. Spangrude, G.I. et al. (1988) Science 241:58.
  3. Spangrude, G.I. et al. (1993) Blood 82:3327.
  4. Morrison, S.J. et al. (1995) Proc. Natl. Acad. Sci. USA 92:10302.
  5. Kawamoto, H. et al. (1997) Int. Immunol. 9:1011.
  6. Yamamoto, Y. et al. (1996) Blood 88:445.
  7. Morrison, S.J. et al. (1997) Proc. Natl. Acad. Sci. USA 94:1908.
  8. Welm, B.E. et al. (2002) Dev. Biol. 245:42.
  9. Codias, E.K. et al. (1990) J. Immunol. 144:2197.
  10. Malek, T.R. et al. (1986) J. Exp. Med. 164:709 .
  11. Codias, E.K. et al. (1990) J. Immunol. 145:1407.
  12. Flood, P.M. et al. (1990) J. Exp. Med. 172:115.

Long Name

Stem Cell Antigen-1/Lymphocyte Antigen 6

Alternate Names

Ly-6, Ly6, Sca1

Entrez Gene IDs

110454 (Mouse)

Gene Symbol

LY6A

UniProt

CAA28351

Additional Sca-1/Ly6 Products

Product Documents for Mouse Sca‑1/Ly6 Antibody

Certificate of Analysis

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Product Specific Notices for Mouse Sca‑1/Ly6 Antibody

For research use only

Citations for Mouse Sca‑1/Ly6 Antibody

Customer Reviews for Mouse Sca‑1/Ly6 Antibody (1)

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  • Mouse Sca-1/Ly6 Antibody
    Name: Adam Guess
    Application: Flow Cytometry
    Sample Tested: bone marrow
    Species: Mouse
    Verified Customer | Posted 11/08/2016
    Mouse Sca‑1/Ly6 Antibody AF1226

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