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BD Pharmingen™ APC-Cy™7 Mouse Anti-Rat CD4
Clone OX-35 (RUO)
Two-color flow cytometric analysis of CD4 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with BD Pharmingen™ FITC Mouse Anti-Rat CD3 antibody (Cat. No. 554832/559975) and either APC-Cy™7 Mouse IgG2a, κ Isotype Control (Cat. No. 565356; Left Panel) or APC-Cy™7 Mouse Anti-Rat CD4 antibody (Cat. No. 565432; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light- scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD4 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with BD Pharmingen™ FITC Mouse Anti-Rat CD3 antibody (Cat. No. 554832/559975) and either APC-Cy™7 Mouse IgG2a, κ Isotype Control (Cat. No. 565356; Left Panel) or APC-Cy™7 Mouse Anti-Rat CD4 antibody (Cat. No. 565432; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light- scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD4 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with BD Pharmingen™ FITC Mouse Anti-Rat CD3 antibody (Cat. No. 554832/559975) and either APC-Cy™7 Mouse IgG2a, κ Isotype Control (Cat. No. 565356; Left Panel) or APC-Cy™7 Mouse Anti-Rat CD4 antibody (Cat. No. 565432; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light- scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD4 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with BD Pharmingen™ FITC Mouse Anti-Rat CD3 antibody (Cat. No. 554832/559975) and either APC-Cy™7 Mouse IgG2a, κ Isotype Control (Cat. No. 565356; Left Panel) or APC-Cy™7 Mouse Anti-Rat CD4 antibody (Cat. No. 565432; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side light- scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
Cd4; CD4 antigen; p55; W3/25 antigen; T-cell surface glycoprotein CD4
Rat (QC Testing)
Mouse BALB/c IgG2a, κ
Rat T-cell blasts
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739236
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO
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 with APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.
Product Notices
- 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.
- 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.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
APC-Cy™7 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
565356
Purified Mouse Anti-Rat CD32 RUO
Size
0.1 mg
Cat No.
550270
Purified Mouse Anti-Rat CD32 RUO
Size
0.5 mg
Cat No.
550271
FITC Mouse Anti-Rat CD3 RUO
Size
0.5 mg
Cat No.
554832
565432 Rev. 2
Antibody Details
OX-35
The OX-35 clone recognizes the CD4 antigen on most thymocytes, a subpopulation of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T helper cells), monocytes, macrophages, some dendritic cells, and microglia. CD4 is an antigen coreceptor on the T-cell surface that interacts with MHC class II molecules on antigen-presenting cells. It participates in T-cell activation through it's association with the T-cell receptor complex and protein tyrosine kinase Lck. The OX-35 clone has been reported to bind to a different epitope of CD4 than that recognized by the W3/25 and OX-38 clones.
565432 Rev. 2
Format Details
APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-Cy7
651 nm
779 nm
565432 Rev.2
Citations & References
View product citations for antibody "565432" on CiteAb
Development References (7)
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Bañuls MP, Alvarez A, Ferrero I, Zapata A, Ardavin C. Cell-surface marker analysis of rat thymic dendritic cells. Immunology. 1993; 79(2):298-304. (Biology). View Reference
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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
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Ford AL, Foulcher E, Goodsall AL, Sedgwick JD. Tissue digestion with dispase substantially reduces lymphocyte and macrophage cell-surface antigen expression. J Immunol Methods. 1996; 194(1):71-75. (Biology: Depletion). View Reference
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Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (Biology). View Reference
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Jefferies WA, Green JR, Williams AF. Authentic T helper CD4 (W3/25) antigen on rat peritoneal macrophages. J Exp Med. 1985; 162(1):117-127. (Immunogen: Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Inhibition). View Reference
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Liu L, Zhang M, Jenkins C, MacPherson GG. Dendritic cell heterogeneity in vivo: two functionally different dendritic cell populations in rat intestinal lymph can be distinguished by CD4 expression. J Immunol. 1998; 161(3):1146-1155. (Biology). View Reference
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Wang CC, Wu CH, Shieh JY, Wen CY, Ling EA. Immunohistochemical study of amoeboid microglial cells in fetal rat brain. J Anat. 1996; 189(3):567-574. (Biology). View Reference
565432 Rev. 2
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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.