Invitrogen

Ly-6A/E (Sca-1) Monoclonal Antibody (D7), PE-eFluor 610, eBioscience™

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Ly-6A/E (Sca-1) Antibody — Figure 3. Reduced frequencies of T cell progenitors in Fermt3 -/- thymi. ( A ) Thymocytes from Fermt3 +/+ and Fermt3 -/- mice were stained for lineage markers (B220, CD19, TER119, NK1.1, CD11b, Gr-1, CD8alpha, CD3e, TCRbeta, TCRgammadelta and CD11c), CD44 and c-kit to identify DN 1-2 (Lin neg , c-kit hi , CD44 hi ) and the DN 3-4 (Lin neg , c-kit low , CD44 low ) populations. ( B and C ) PB from Fermt3 +/+ and Fermt3 -/- animals ( P3 ) were stained for lineage markers (TER119, B220, CD11b, Gr-1, CD11c, NK1.1, CD4 and CD8alpha), c-kit and Sca-1 to identify hematopoietic progenitor cells and analysed by flow cytometry. N = 4. Numbers within the representative FACS plots indicate cell percentages. Bars indicate means +- standard errors. **p < 0.01; ***p < 0.001. See also Figure 3--figure supplement 1 . Figure 3--figure supplement 1. Reduced frequencies of T cell prognitors in Fermt3 -/- thymi. Thymocytes from Fermt3 +/+ and Fermt3 -/- P3 mice were stained for lineage markers (B220, CD19, TER119, NK1.1, CD11b, Gr-1, CD8alpha, CD3epsilon, TCRbeta, TCRgammadelta and CD11c), CD44 and CD25 to identify and quantify DN1 (Lin neg , CD44 hi , CD25 - ), DN2 (Lin neg , CD44 hi , CD25 + ), DN3 (Lin neg , CD44 low , CD25 + ) and DN4 (Lin neg , CD44 low , CD25 - ) populations by flow cytometry. Numbers within FACS plots indicate cell percentages. Total number of DN1, DN2, DN3 and DN4 cells were measured by flow cytometry. N = 20. Bars indicate means +- stand

Ly-6A/E (Sca-1) Antibody in Flow Cytometry (Flow) — Staining of C57Bl/6 bone marrow cells with Mouse Hematopoietic Lineage Cocktail eFluor® 450 (Product # 88-7772-72), Anti-Mouse CD117 (c-Kit) APC (Product # 17-1171-82), and 0.25 µg of Rat IgG2a K Isotype Control PE-eFluor® 610 (Product # 61-4321-82) (left) or 0.25 µg of Anti-Mouse Ly-6A/E (Sca-1) PE-eFluor® 610 (right). Lineage negative/low viable cells, as determined by Fixable Viability Dye eFluor® 506 (Product # 65-0866-14), were used for analysis.

Product Details

Tested Applications

Dilution

Flow Cytometry (Flow)

0.5 µg/test

Published Applications

Flow Cytometry (Flow)

See 28 publications below

Product Specifications

Species Reactivity

Mouse

Published species

Fish

Host / Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), PE-eFluor 610, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

Conjugate

PE-eFluor® 610

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

RRID

AB_2574648

Product Specific Information

Description: The D7 monoclonal antibody reacts with mouse Sca-1, an 18 kDa member of the Ly-6 family of GPI-linked surface proteins. D7 reacts with both Ly-6E.1 and Ly-6A.2 molecules expressed by mouse hematopoietic stem cells, myeloid population and peripheral T and B cells. Non-activated splenocytes from Ly-6.2 strains including C57BL, SJL, 129, AKR, and others demonstrate higher frequency of D7-positive cells compared to the Ly-6.1 strains such as BALB/c, C3H, NZB and others. Sca-1 is expressed at high levels upon activation regardless of the Ly-6 haplotype. The Ly-6 family is involved in regulation and function of T cell activation. Sca-1 is a major phenotypic marker for mouse hematopoietic progenitor/stem cell subset.

Applications Reported: This D7 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This D7 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PE-eFluor® 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red®). Please make sure that your instrument is capable of detecting this fluorochome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488-561 nm; Emission: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Ly-6A/E molecules are implicated in the regulation of T cell activation.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: locus A/E; Ly-6A.2/Ly-6E.1; Ly-6E; lymphocyte antigen 6 complex; Lymphocyte antigen 6A-2/6E-1; Lymphocyte antigen 6E; SCA-1; Stem cell antigen 1; Stem cell antigen 2; T-cell-activating protein; TAP; Thymic shared antigen 1; TSA-1

Gene Aliases: 9804; Ly-6A.2; Ly-6A/E; Ly-6E.1; Ly6; Ly67; Ly6a; Ly6e; RIG-E; Sca-1; Sca-2; Sca1; TAP; TSA-1; Tsa1

UniProt ID: (Mouse) P05533, (Mouse) Q64253

Entrez Gene ID: (Mouse) 110454, (Mouse) 17069

Function(s)

in utero embryonic development adrenal gland development organ growth norepinephrine metabolic process epinephrine secretion ventricular cardiac muscle tissue morphogenesis

Ly-6A/E (Sca-1) Monoclonal Antibody (D7), PE-eFluor 610, eBioscience™

Product Details

Product Specific Information

Target Information

Bioinformatics

Material safety data sheets (MSDS)

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