Flow cytometric analysis of CD8 expression on pig peripheral blood lymphocytes. Pig whole blood was stained simultaneously with PE Mouse Anti-Pig CD4a antibody (Cat. No. 559586) and Alexa Fluor® 647 Mouse Anti-Pig CD8a antibody (Cat. no. 561475). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plot shows the expression of CD8 versus CD4 derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD8 expression on pig peripheral blood lymphocytes. Pig whole blood was stained simultaneously with PE Mouse Anti-Pig CD4a antibody (Cat. No. 559586) and Alexa Fluor® 647 Mouse Anti-Pig CD8a antibody (Cat. no. 561475). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The two-color flow cytometric dot plot shows the expression of CD8 versus CD4 derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
Pig (QC Testing)
Mouse BALB/c IgG2a, κ
dd miniature swine thymocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_10713174
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE Mouse Anti-Pig CD4a RUO
Size
0.1 mg
Cat No.
559586
561475 Rev. 1
Antibody Details
76-2-11
The 76-2-11 (also known as clone PT8) monoclonal antibody specifically binds to an epitope on the CD8α chain, a 35-kDa antigen expressed on thymocytes, peripheral T lymphocytes, and NK cells. The CD8 molecule can exist as a 70 kDa homodimer, composed of α chains, or heterodimer, composed of an α and β chain. Cells which express the CD8αα homodimer display dimmer staining with mAb 76-2-11 than CDαβ-expressing cells. The 76-2-11 mAb does not cross-react with human or bovine cells. Two peripheral CD8+ T-cell populations can be distinguished in the pig: CD8-bright CD4- CTL effectors/precursors and CD8-dull CD4+ T-helper lymphocytes. Pig NK cells express CD8 (dull staining), CD2, MHC class II, LFA-1, and asialo-GM1, but not CD3, CD4, CD5, or CD6. mAb 76-2-11 has been reported to partially inhibit in vitro cytotoxic activity of PBL to allogeneic leukocytes, but not NK-cell-mediated lysis, and to deplete CD8+ T cells in vivo. This clone was clustered as anti-CD8a at the First International Swine CD workshop.
561475 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
561475 Rev.1
Citations & References
Development References (13)
-
Dato ME, Kim YB. Characterization and utilization of a monoclonal antibody inhibiting porcine natural killer cell activity for isolation of natural killer and killer cells. J Immunol. 1990; 144(11):4452-4462. (Biology). View Reference
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Pescovitz MD, Lowman MA, Sachs DH. Expression of T-cell associated antigens by porcine natural killer cells. Immunology. 1988; 65(2):267-271. (Biology). View Reference
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Pescovitz MD, Lunney JK, Sachs DH. Murine anti-swine T4 and T8 monoclonal antibodies: distribution and effects on proliferative and cytotoxic T cells. J Immunol. 1985; 134(1):37-44. (Biology). View Reference
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Pescovitz MD, Lunney JK, Sachs DH. Preparation and characterization of monoclonal antibodies reactive with porcine PBL. J Immunol. 1984; 133(1):368-375. (Immunogen). View Reference
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Saalmuller A, Aasted B, Canals A, et al. Analyses of mAb reactive with porcine CD8. Vet Immunol Immunopathol. 1994; 43(1-3):249-254. (Clone-specific). View Reference
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Saalmuller A, Hirt W, Maurer S, Weiland E. Discrimination between two subsets of porcine CD8+ cytolytic T lymphocytes by the expression of CD5 antigen. Immunology. 1994; 81(4):578-583. (Biology). View Reference
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Saalmuller A, Pauly T, Hohlich BJ, Pfaff E. Characterization of porcine T lymphocytes and their immune response against viral antigens. J Biotechnol. 1999; 73(2-3):223-233. (Biology). View Reference
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Saalmuller A, Werner T, Fachinger V. T-helper cells from naive to committed. Vet Immunol Immunopathol. 2002; 87(3-4):137-145. (Biology). View Reference
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Smith CV, Sablinski T, Arn JS, et al. In vivo treatment with monoclonal antibodies directed against CD4 and CD8 antigens in miniature swine. J Immunother Emphasis Tumor Immunol. 1994; 16(2):105-114. (Biology). View Reference
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Suzuki T, Sundt TM 3rd, Mixon A, Sachs DH. In vivo treatment with antiporcine T cell antibodies. Transplantation. 1990; 50(1):76-81. (Biology). View Reference
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Waters WR, Hontecillas R, Sacco RE, et al. Antigen-specific proliferation of porcine CD8alphaalpha cells to an extracellular bacterial pathogen. Immunology. 2000; 101(3):333-341. (Clone-specific). View Reference
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Zuckermann FA, Pescovitz MD, Aasted B, et al. Report on the analyses of mAb reactive with porcine CD8 for the second international swine CD workshop. Vet Immunol Immunopathol. 1998; 60(3-4):291-303. (Clone-specific). View Reference
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Zuckermann FA. Extrathymic CD4/CD8 double positive T cells. Vet Immunol Immunopathol. 1999; 72(1-2):55-66. (Biology). View Reference
561475 Rev. 1
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.
