PE-CF594 Rat Anti-Mouse Ly-6C

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BD Horizon™ PE-CF594 Rat Anti-Mouse Ly-6C

Name: PE-CF594 Rat Anti-Mouse Ly-6C
Brand: PE-CF594 Rat Anti-Mouse Ly-6C
SKU: 570134

Clone HK1.4.rMAb (also known as HK1.4) (RUO)

Multicolor flow cytometric analysis of Ly-6C expression on Mouse splenic leukocytes. Top Plots: C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with APC Rat Anti-Mouse CD8a antibody (Cat. No. 553035) and with either BD Horizon™ PE-CF594 Rat IgG1, κ Isotype Control (Cat. No. 562309; Left Plot) or BD Horizon™ PE-CF594 Rat Anti-Mouse Ly-6C antibody (Cat. No. 570053/570134; Right Plot) at 0.5 μg/test. Bottom Plots: C57BL/6 Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™). The cells were then stained with BD Horizon™ BB515 Rat Anti-Mouse CD11b antibody (Cat. No. 564454) and with either BD Horizon™ PE-CF594 Rat IgG1, κ Isotype Control (Left Plot) or BD Horizon™ PE-CF594 Rat Anti-Mouse Ly-6C antibody (Right Plot) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of Ly-6C (or Ig Isotype control staining) versus CD8a (Top Plots) or CD11b (Bottom Plots) were derived from gated events with the forward and side-light scatter characteristics of viable (DAPI-negative) leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Horizon™

Alternative Name: Ly-6C; Ly-6C1; Ly6c; Ly6c1; Lymphocyte antigen Ly-6C

Reactivity: Mouse (QC Testing)

Isotype: Rat IgG1, κ

Immunogen: L3 Clone CTL Cells

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Entrez Gene ID: 17067

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.
Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
CF™ is a trademark of Biotium, Inc.
When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

Size 0.5 mg Cat No. 553142

APC Rat Anti-Mouse CD8a RUO

Size 0.1 mg Cat No. 553035

PE-CF594 Rat IgG1, κ Isotype Control RUO

Size 0.1 mg Cat No. 562309

View Details

570134 Rev. 1

Antibody Details

HK1.4.rMAb

The HK1.4.rMAb monoclonal antibody is a recombinant monoclonal antibody derived from HK1.4 hybridoma cells that specifically recognizes a non-polymorphic determinant on Mouse Lymphocyte antigen Ly-6C which is also known as Ly6c. Ly-6C is an ~14-17 kDa glycosylphosphatidylinositol (GPI)-linked cell-surface antigen that is encoded by Ly6c1 (Lymphocyte antigen 6 complex, locus C1) which belongs to the Ly-6 gene family. Ly-6C is expressed on monocytes, macrophages, neutrophils, eosinophils, endothelial cells, plasma cells, thymocytes, NK cells, and some T cell subsets. Mice with the Ly-6.2 haplotype (eg, AKR, C57BL, C57BR, C57L, C58, DBA/2, PL, SJL, SWR, 129) have subsets of CD8+ and CD4+ Ly-6C+ T cells, while Ly-6.1 strains (eg, A, BALB/c, CBA, C3H/He, DBA/1, NZB) have only CD8+ Ly-6C+ T cells. Upregulation of Ly-6C expression on CD8+ T cells by interferons α and β and poly (I:C) has been described, and Ly-6C is a memory marker on CD8+ T cells. The HK1.4 antibody does not reportedly block the binding of the RB6-8C5 monoclonal antibody that specifically recognizes Mouse Ly-6G and Ly-6C.

570134 Rev. 1

Format Details

PE-CF594

BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE-CF594

Excitation Source: Yellow-Green 561 nm

Excitation Max: 496 nm, 566 nm

Emission Max: 615 nm

570134 Rev.1

Citations & References

View product citations for antibody "570134" on CiteAb

Development References (3)

Havran WL, Lancki DW, Moldwin RL, Dialynas DP, Fitch FW. Characterization of an anti-Ly-6 monoclonal antibody which defines and activates cytolytic T lymphocytes.. J Immunol. 1988; 140(4):1034-42. (Immunogen: Activation, Blocking, (Co)-stimulation, Flow cytometry, Fluorescence activated cell sorting, Functional assay, Stimulation). View Reference
Ma C, Kapanadze T, Gamrekelashvili J, Manns MP, Korangy F, Greten TF. Anti-Gr-1 antibody depletion fails to eliminate hepatic myeloid-derived suppressor cells in tumor-bearing mice.. J Leukoc Biol. 2012; 92(6):1199-206. (Clone-specific: Flow cytometry). View Reference
Schlueter AJ, Malek TR, Hostetler CN, Smith PA, deVries P, Waldschmidt TJ. Distribution of Ly-6C on lymphocyte subsets: I. Influence of allotype on T lymphocyte expression.. J Immunol. 1997; 158(9):4211-22. (Biology). View Reference

570134 Rev. 1

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.