Sca-1/Ly6 Antibody [Unconjugated]

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.

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 (//pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//dx.plos.org/10.1371/journal.pone.0113247), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//dx.plos.org/10.1371/journal.pone.0113247), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/19344510), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, Flow, CyTOF-ready, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Sca-1/Ly6 Antibody [Unconjugated] Summary

• 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: Ly6a
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Sca-1/Ly6 Antibody [Unconjugated]

• Ly6
• Ly-6
• Ly-6A.2
• Ly-6A/E
• Ly-6E.1
• lymphocyte antigen 6 complex, locus A
• lymphocyte antigen 6A-2/6E-1
• Sca1
• Sca-1
• Stem cell antigen 1
• TAP
• T-cell-activating protein

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Sca-1/Ly6 Antibody (AF1226)(30)

Publications using AF1226

• Skulimowska, I;Morys, J;Sosniak, J;Gonka, M;Gulati, G;Sinha, R;Kowalski, K;Mosiolek, S;Weissman, IL;Jozkowicz, A;Szade, A;Szade, K; Polyclonal regeneration of mouse bone marrow endothelial cells after irradiative conditioning Cell reports 2024-10-25 [PMID: 39489938] (Immunohistochemistry, Mouse, Transgenic Mouse)
• Lee, JY;Reyes, N;Woo, SH;Goel, S;Stratton, F;Kuang, C;Mansfield, AS;LaFave, LM;Peng, T; Senescent fibroblasts in the tumor stroma rewire lung cancer metabolism and plasticity bioRxiv : the preprint server for biology 2024-07-30 [PMID: 39131266] (Immunohistochemistry, Transgenic Mouse)
• EC Jeffery, TLA Mann, JA Pool, Z Zhao, SJ Morrison Bone marrow and periosteal skeletal stem/progenitor cells make distinct contributions to bone maintenance and repair Cell Stem Cell, 2022-10-21;29(11):1547-1561.e6. 2022-10-21 [PMID: 36272401] (IHC, Mouse)
• H Cui, Z Li, S Chen, X Li, D Chen, J Wang, Z Li, W Hao, F Zhong, K Zhang, Z Zheng, Z Zhan, H Liu CXCL12/CXCR4-Rac1-mediated migration of osteogenic precursor cells contributes to pathological new bone formation in ankylosing spondylitis Science Advances, 2022-04-06;8(14):eabl8054. 2022-04-06 [PMID: 35385310] (IHC, Mouse, Transgenic Mouse)
• F Kleefeldt, B Upcin, H Bömmel, C Schulz, G Eckner, J Allmanritt, J Bauer, B Braunger, U Rueckschlo, S Ergün Bone marrow-independent adventitial macrophage progenitor cells contribute to angiogenesis Cell Death & Disease, 2022-03-09;13(3):220. 2022-03-09 [PMID: 35264563] (ICC, Mouse)
• ES Kim, BH Yoon, SM Lee, M Choi, EH Kim, BW Lee, SY Kim, CG Pack, YH Sung, IJ Baek, CH Jung, TB Kim, JY Jeong, CH Ha Fecal microbiota transplantation ameliorates atherosclerosis in mice with C1q/TNF-related protein 9 genetic deficiency Experimental & Molecular Medicine, 2022-02-03;0(0):. 2022-02-03 [PMID: 35115674] (IHC, Mouse)
• P Pagella, C Nombela-Ar, TA Mitsiadis Distinct Expression Patterns of Cxcl12 in Mesenchymal Stem Cell Niches of Intact and Injured Rodent Teeth International Journal of Molecular Sciences, 2021-03-16;22(6):. 2021-03-16 [PMID: 33809663] (IHC, Mouse)
• Philipp S. Lienemann, Queralt Vallmajo‐Martin, Panagiota Papageorgiou, Ulrich Blache, Stéphanie Metzger, Anna‐Sofia Kiveliö et al. Smart Hydrogels for the Augmentation of Bone Regeneration by Endogenous Mesenchymal Progenitor Cell Recruitment Advanced Science 4/1/2020 [PMID: 32274319]
• Philipp S. Lienemann, Queralt Vallmajo‐Martin, Panagiota Papageorgiou, Ulrich Blache, Stéphanie Metzger, Anna‐Sofia Kiveliö et al. Smart Hydrogels for the Augmentation of Bone Regeneration by Endogenous Mesenchymal Progenitor Cell Recruitment Advanced Science 2020-04-01 [PMID: 32274319] (Immunohistochemistry, Mouse)
• Tan GK, Pryce BA, Stabio A et al. Tgfbeta signaling is critical for maintenance of the tendon cell fate Elife 1/22/2020 [PMID: 31961320]
• Helen L. Moser, Anton Ph. Doe, Kristen Meier, Simon Garnier, Damien Laudier, Haruhiko Akiyama et al. Genetic lineage tracing of targeted cell populations during enthesis healing Journal of Orthopaedic Research® 12/1/2018 [PMID: 30084210]
• DL Coutu, KD Kokkaliari, L Kunz, T Schroeder Multicolor quantitative confocal imaging cytometry Nat. Methods, 2017-11-13;15(1):39-46. 2017-11-13 [PMID: 29320487] (IHC, Mouse)
• L Golle, HU Gerth, K Beul, B Heitplatz, P Barth, M Fobker, H Pavenstädt, GS Di Marco, M Brand Bone marrow-derived cells and their conditioned medium induce microvascular repair in uremic rats by stimulation of endogenous repair mechanisms Sci Rep, 2017-08-25;7(1):9444. 2017-08-25 [PMID: 28842629] (Flow Cytometry, Rat)
• E Zhu, L Hu, H Wu, L Piao, G Zhao, A Inoue, W Kim, C Yu, W Xu, YK Bando, X Li, Y Lei, CN Hao, K Takeshita, WS Kim, K Okumura, T Murohara, M Kuzuya, XW Cheng Dipeptidyl Peptidase-4 Regulates Hematopoietic Stem Cell Activation in Response to Chronic Stress J Am Heart Assoc, 2017-07-14;6(7):. 2017-07-14 [PMID: 28710180] (ICC, Mouse)
• JM Daniel, J Dutzmann, H Brunsch, J Bauersachs, R Braun-Dull, DG Sedding Systemic application of sirolimus prevents neointima formation not via a direct anti-proliferative effect but via its anti-inflammatory properties Int. J. Cardiol., 2017-03-14;0(0):. 2017-03-14 [PMID: 28433555] (IHC, Mouse)
• Beate M. Lichtenberger, Maria Mastrogiannaki, Fiona M. Watt Epidermal beta -catenin activation remodels the dermis via paracrine signalling to distinct fibroblast lineages Nature Communications 2/3/2016 [PMID: 26837596]
• Stephanie L. Easter, Elizabeth H. Mitchell, Sarah E. Baxley, Renee Desmond, Andra R. Frost, Rosa Serra Wnt5a Suppresses Tumor Formation and Redirects Tumor Phenotype in MMTV-Wnt1 Tumors PLoS ONE 11/17/2014 [PMID: 25401739]
• Jumabay M, Abdmaulen R, Ly A, Cubberly M, Shahmirian L, Heydarkhan-Hagvall S, Dumesic D, Yao Y, Bostrom K Pluripotent stem cells derived from mouse and human white mature adipocytes. Stem Cells Transl Med, 2014-01-06;3(2):161-71. 2014-01-06 [PMID: 24396033] (ICC, Mouse)
• Driskell R, Lichtenberger B, Hoste E, Kretzschmar K, Simons B, Charalambous M, Ferron S, Herault Y, Pavlovic G, Ferguson-Smith A, Watt F Distinct fibroblast lineages determine dermal architecture in skin development and repair. Nature, 2013-12-12;504(7479):277-81. 2013-12-12 [PMID: 24336287] (IHC-Fr, Mouse)
• Paxson J, Gruntman A, Davis A, Parkin C, Ingenito E, Hoffman A Age dependence of lung mesenchymal stromal cell dynamics following pneumonectomy. Stem Cells Dev, 2013-08-30;22(24):3214-25. 2013-08-30 [PMID: 23895415] (IHC-P, Mouse)
• Fujio M, Yamamoto A, Ando Y, Shohara R, Kinoshita K, Kaneko T, Hibi H, Ueda M Stromal cell-derived factor-1 enhances distraction osteogenesis-mediated skeletal tissue regeneration through the recruitment of endothelial precursors. Bone, 2011-06-29;49(4):693-700. 2011-06-29 [PMID: 21741502] (IHC-Fr, Mouse)
• Fujita-Sato S, Ito S, Isobe T, Ohyama T, Wakabayashi K, Morishita K, Ando O, Isono F Structural basis of digoxin that antagonizes RORgamma t receptor activity and suppresses Th17 cell differentiation and interleukin (IL)-17 production. J. Biol. Chem., 2011-07-06;286(36):31409-17. 2011-07-06 [PMID: 21733845] (IHC-Fr, Mouse)
• Smith BH, Gazda LS, Conn BL Three-dimensional culture of mouse renal carcinoma cells in agarose macrobeads selects for a subpopulation of cells with cancer stem cell or cancer progenitor properties. Cancer Res., 2011-01-24;71(3):716-24. 2011-01-24 [PMID: 21266363] (IHC, Mouse)
• Yoshinori Sumita, Younan Liu, Saeed Khalili, Ola M. Maria, Dengsheng Xia, Sharon Key et al. Bone marrow-derived cells rescue salivary gland function in mice with head and neck irradiation The International Journal of Biochemistry & Cell Biology 1/1/2011 [PMID: 20933096]
• Germano D, Blyszczuk P, Valaperti A, Kania G, Dirnhofer S, Landmesser U, Luscher TF, Hunziker L, Zulewski H, Eriksson U Prominin-1/CD133+ lung epithelial progenitors protect from bleomycin-induced pulmonary fibrosis. Am. J. Respir. Crit. Care Med., 2009-02-20;179(10):939-49. 2009-02-20 [PMID: 19234103] (Flow Cytometry, Mouse)
• Zhang Y, Goss AM, Cohen ED, Kadzik R, Lepore JJ, Muthukumaraswamy K, Yang J, DeMayo FJ, Whitsett JA, Parmacek MS, Morrisey EE A Gata6-Wnt pathway required for epithelial stem cell development and airway regeneration. Nat. Genet., 2008-06-08;40(7):862-70. 2008-06-08 [PMID: 18536717] (ICC, Mouse)
• Flanagan K, Modrusan Z, Cornelius J, Chavali A, Kasman I, Komuves L, Mo L, Diehl L Intestinal epithelial cell up-regulation of LY6 molecules during colitis results in enhanced chemokine secretion. J. Immunol., 2008-03-15;180(6):3874-81. 2008-03-15 [PMID: 18322195] (IHC-Fr, Mouse)
• Vranken I, De Visscher G, Lebacq A, Verbeken E, Flameng W The recruitment of primitive Lin(-) Sca-1(+), CD34(+), c-kit(+) and CD271(+) cells during the early intraperitoneal foreign body reaction. Biomaterials, 2007-11-26;29(7):797-808. 2007-11-26 [PMID: 18022690] (ICC, Rat)
• Ling TY, Kuo MD, Li CL, Yu AL, Huang YH, Wu TJ, Lin YC, Chen SH, Yu J Identification of pulmonary Oct-4+ stem/progenitor cells and demonstration of their susceptibility to SARS coronavirus (SARS-CoV) infection in vitro. Proc. Natl. Acad. Sci. U.S.A., 2006-06-13;103(25):9530-5. 2006-06-13 [PMID: 16772384] (ICC, Mouse)
• Kim CF, Jackson EL, Woolfenden AE, Lawrence S, Babar I, Vogel S, Crowley D, Bronson RT, Jacks T Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell, 2005-06-17;121(6):823-35. 2005-06-17 [PMID: 15960971] (ICC, Mouse)

Bioinformatics

• Gene Symbol: Ly6a
• Uniprot:
  • CAA28351()