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Purified Rat Anti-Human CD132
Purified Rat Anti-Human CD132
Flow cytometric analysis of CD132 expression on human peripheral blood lymphocytes. Whole blood was stained with either Purified Rat IgG2b, κ Isotype Control (Cat. No. 555846; dashed line histogram) or Purified Rat Anti-Human CD132 (Cat. No. 555896; solid line histogram), followed with Biotin Goat Anti-Rat Ig (Cat. No. 554014) and PE Streptavidin (Cat. No. 555896). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 349202). Fluorescent histograms depicting CD132 (or Ig isotype) expression were from event with the forward and side light-scattering characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACScan™ system.
Flow cytometric analysis of CD132 expression on human peripheral blood lymphocytes. Whole blood was stained with either Purified Rat IgG2b, κ Isotype Control (Cat. No. 555846; dashed line histogram) or Purified Rat Anti-Human CD132 (Cat. No. 555896; solid line histogram), followed with Biotin Goat Anti-Rat Ig (Cat. No. 554014) and PE Streptavidin (Cat. No. 555896). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 349202). Fluorescent histograms depicting CD132 (or Ig isotype) expression were from event with the forward and side light-scattering characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACScan™ system.
Product Details
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BD Pharmingen™
IL-2RG; IL-2Rγ; γc; Common γ chain; Common gamma chain; gamma c; SCIDX1
Human (QC Testing), Dog (Tested in Development)
Rat IgG2b, κ
Human γc Transfected Cell Line
Flow cytometry (Routinely Tested)
0.5 mg/ml
VI C-89
AB_396208
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

This reagent is effective for indirect immunofluorescence staining of human tissue for flow cytometric analysis. For flow cytometric applications, a three step labelling procedure is recommended for amplifying signal.

Suggested protocol for 3-step staining using Lysed Whole Blood method:

1. Incubate 100 µl whole blood with primary (unconjugated) antibody for 20-30 minutes at room temperature.

2. Add 2 mls of 1X Pharm Lyse (10X Pharm Lyse, Cat. No. 555899) and incubate for 10-15 minutes. Centrifuge and aspirate.

3. Wash once with PBS/0.1% sodium azide/ 1% heat-inactivated fetal bovine serum (PBS-FBS). Centrifuge and aspirate.

4. Add biotinylated mouse anti-rat Ig's (Cat. No. 553883) and incubate for 20-30 minutes at room temperature.

5. Wash once with PBS-FBS. Centrifuge and aspirate.

6. Add SAV-PE (Cat. No. 554061) and incubate for 20-30 minutes in the dark at room temperature.

7. Wash once with PBS-FBS. Centrifuge and aspirate Resuspend in 0.5 ml of PBS-FBS and analyze by flow cytometry.

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. 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.
  4. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
555896 Rev. 10
Antibody Details
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TUGh4

The TUGh4 monoclonal antibody specifically binds to CD132. CD132 is a 65-70 kDa type 1 transmembrane glycoprotein that is encoded by the IL2RG (interleukin 2 receptor, gamma) gene. CD132 is also known as the common γ subunit (γc) and is shared by the IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21 receptor complexes. CD132 is broadly expressed by most peripheral T and B lymphocytes, NK cells, monocytes, and granulocytes. The cytoplasmic domain of the γc chain plays an important role in cytokine-mediated signal transduction. Mutation of the IL2RG gene results in X-linked severe combined immunodeficiency (XSCID). The TUGh4 antibody recognizes a different epitope from that recognized by the CD132-specific clone, AG184.

555896 Rev. 10
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
555896 Rev.10
Citations & References
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Development References (5)

  1. Ishii N, Kondo M, Takeshita T, and Sugamura K. mAb specific for the γ chain of the IL-2 receptor. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:1867-1868.
  2. Ishii N, Takeshita T, Kimura Y, et al. Expression of the IL-2 receptor gamma chain on various populations in human peripheral blood. Int Immunol. 1994; 6(8):1273-1277. (Biology). View Reference
  3. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  4. Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
  5. Matsuoka M, Takeshita T, Ishii N, Nakamura M, Ohkubo T, Sugamura K. Kinetic study of interleukin-2 binding on the reconstituted interleukin-2 receptor complexes including the human gamma chain. Eur J Immunol. 1993; 23(10):2472-2476. (Biology). View Reference
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555896 Rev. 10

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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.