Dog T Lymphocyte Cocktail

BD Pharmingen™ Dog T Lymphocyte Cocktail

Name: Dog T Lymphocyte Cocktail
Brand: BD Pharmingen™
SKU: 558699

(RUO)

Protocol Library

Scientific Resources

Three-color analysis of the expression of CD3, CD4, and CD8 on lysed canine whole blood. Dog PBMCs were stained with either Isotype Control Cocktail A (Cat. No. 558509; Panel D) or Dog T Lymphocyte Cocktail (Cat. No. 558699). During data analysis, lymphocytes were identified by scatter profile. Panel A represents the Pan-T and CD4 profile, panel B represent the Pan-T with CD8 profile while panel C shows CD4 and CD8 profile. Flow cytometry was performed on a BD FACSCalibur™.

Protocol Library

Scientific Resources

Product Details

Brand: BD Pharmingen™

Reactivity: Dog (QC Testing)

Application: Flow cytometry (Routinely Tested)

Regulatory Status: RUO

RRID: AB_1645609

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

Description

The Dog T Lymphocyte Cocktail is a three-color reagent cocktail designed to identify dog T lymphocytes by direct immunofluorescence staining with flow cytometric analysis. The LSM 1.140 antibody reacts with a CD8-like 32-35 kDa heterodimer. The CD8α and β chains (CD8a and CD8b, respectively) form a heterodimer on the surface of most thymocytes and a subpopuplation of mature T-lymphocytes (ie, MHC class I-restricted T cells, including most T suppressor/cytotoxic cells). The LSM 12.125 antibody reacts with the canine form of the 56 kDa transmembrane glycoprotein, CD4, present on the T-helper/inducer subset of normal canine donor peripheral blood lymphocytes. The distribution on lymphocytes for canine is similar to that found for both human and monkey, with the majority of CD4-positive lymphocytes being CD8-negative and lacking reactivity with antibodies to B- or NK-cell markers. CD4 in the canine is also present on monocytes and granulocytes, but in higher numbers than what is seen for human. The LSM 8.358 antibody reacts with a CD3-like T-cell receptor-associated cell-surface antigen found on thymocytes and peripheral T lymphocytes. These monoclonals were generated by immunizing mice with freshly isolated canine thymocytes.

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
An isotype control should be used at the same concentration as the antibody of interest.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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.
This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

558699 Rev. 3

Components

Description	Clone	Isotype	EntrezGene ID
APC Mouse anti-Dog Pan T Cell	LSM 8.358.1.1	IgM, κ	N/A
PE Mouse anti-Dog CD4 (IL-2R alpha chain)	LSM 12.125	IgG1,	N/A
FITC anti-Dog CD8	LSM 1.140	IgG1, κ	N/A

558699 Rev. 3

Citations & References

Development References (4)

Bierer BE, Sleckman BP, Ratnofsky SE, Burakoff SJ. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989; 7:579-599. (Biology). View Reference
Cobbold S, Metcalfe S. Monoclonal antibodies that define canine homologues of human CD antigens: Summary of the First International Canine Leukocyte Antigen Workshop (CLAW). Tissue Antigens. 1994; 43:137-154. (Clone-specific). View Reference
Gebhard GH, Carter PB. Identification of canine T-lymphocyte subsets with monoclonal antibodies. Vet Immunol Immunopathol. 1992; 33:187-199. (Immunogen). View Reference
Ruslander DA, Gebhard DH, Tompkins MB, Grindem CB, Page RL. Immunophenotypic characterization of canine lymphoproliferative disorders. In Vivo. 1997; 11(2):169-172. (Biology). View Reference

558699 Rev. 3

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

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.