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Alexa Fluor® 647 Mouse Anti-Pig CD4a
Alexa Fluor® 647 Mouse Anti-Pig CD4a
Multicolor flow cytometric analysis of CD4 expression on pig peripheral blood lymphocytes. Pig whole blood was stained simultaneously with Alexa Fluor® 647 Mouse Anti-Pig CD4a antibody (Cat. No. 561472) and PE Mouse Anti-Pig CD3ε antibody (Cat. No. 561485). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric dot plots showing the correlated expression of CD4a versus CD3 were 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.
Multicolor flow cytometric analysis of CD4 expression on pig peripheral blood lymphocytes. Pig whole blood was stained simultaneously with Alexa Fluor® 647 Mouse Anti-Pig CD4a antibody (Cat. No. 561472) and PE Mouse Anti-Pig CD3ε antibody (Cat. No. 561485). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). Two-color flow cytometric dot plots showing the correlated expression of CD4a versus CD3 were 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
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BD Pharmingen™
CD4; CD4 molecule; Lymphocyte antigen CD4
Pig (QC Testing)
Mouse BALB/c IgG2b, κ
dd miniature swine thymocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_10680272
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

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. 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.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
561472 Rev. 1
Antibody Details
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74-12-4

The 74-12-4 (also known as clone PT4) monoclonal antibody specifically binds to CD4, a 55-kDa antigen expressed on T lymphocytes. This antibody does not react with CTL effectors, CTL precursors, or NK cells (ie, CD8[bright] cells) and it does not cross-react with human or bovine cells. Two peripheral T-helper lymphocyte phenotypes can be distinguished in the pig: CD4+CD8- and CD4+CD8[dull]. mAb 74-12-4 has been reported to inhibit proliferative responses of peripheral blood lymphocytes to mitogen, soluble antigen, and alloantigen. It is only marginally effective for in vivo depletion of peripheral CD4+ T cells. Two alloantigenic forms of CD4 have been recognized in miniature swine based upon their recognition (CD4.1) or lack of recognition (CD4.2) by mAb 74-12-4; the CD4.2 phenotype displays an autosomal recessive, non-MHC-linked, pattern of inheritance.  The molecular basis for the polymorphism is a cluster of  nucleotide differences leading to multiple amino-acid substitutions in the Ig CDR2-like loop structure. This mAb was clustered as anti-CD4a at the First International Swine CD Workshop.  It has been reported to crossreact with chicken leukocytes.

561472 Rev. 1
Format Details
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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
561472 Rev.1
Citations & References
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Development References (10)

  1. Brodersen R, Bijlsma F, Gori K. Analysis of the immunological cross reactivities of 213 well characterized monoclonal antibodies with specificities against various leucocyte surface antigens of human and 11 animal species. Vet Immunol Immunopathol. 1998; 64(1):1-13. (Biology). View Reference
  2. 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. (Immunogen). View Reference
  3. Gustafsson K, Germana S, Sundt TM, Sachs DH, LeGuern C. Extensive allelic polymorphism in the CDR2-like region of the miniature swine CD4 molecule. J Immunol. 1993; 151(3):1365-1370. (Biology). View Reference
  4. 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
  5. Pescovitz MD, Lunney JK, Sachs DH. Preparation and characterization of monoclonal antibodies reactive with porcine PBL. J Immunol. 1984; 133(1):368-375. (Biology). View Reference
  6. Saalmuller A, Aasted B, Canals A, et al. Analyses of mAb reactive with porcine CD8. Vet Immunol Immunopathol. 1994; 43(1-3):249-254. (Biology). View Reference
  7. Saalmüller A, Aasted B, Canals A. Summary of workshop findings for porcine T-lymphocyte antigens. Vet Immunol Immunopathol. 1994; 43(1-3):219-228. (Biology). View Reference
  8. 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
  9. Sundt TM, LeGuern C, Germana S. Characterization of a polymorphism of CD4 in miniature swine. J Immunol. 1992; 148(10):3195-3201. (Biology). View Reference
  10. 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
View All (10) View Less
561472 Rev. 1

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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.