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65991A_556434_image1.png

Flow cytometric analysis of perforin expression in human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Purified Mouse IgG2b, κ Isotype Control (Dashed line histogram; Cat No. 555740/556654) or Purified Mouse Anti-Human Perforin (Cat. No 556434), followed by a FITC conjugated secondary antibody. Flow cytometric analysis was performed using a BD FACScan™  Flow Cytometer System.

65991A_556434_image2.png

Acetone-fixed, frozen tissue section of human lymphoma stained with Purified Mouse Anti-Human Perforin  (Cat. No. 556434) using a DAB chromogen and Hematoxylin counterstain. Staining is seen in scattered infiltrated lymphocytes, arrows indicate examples.

65991A_556434_image1.png 65991A_556434_image2.png Blue-Cap-White-Label.jpg

Flow cytometric analysis of perforin expression in human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were fixed and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed with and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Purified Mouse IgG2b, κ Isotype Control (Dashed line histogram; Cat No. 555740/556654) or Purified Mouse Anti-Human Perforin (Cat. No 556434), followed by a FITC conjugated secondary antibody. Flow cytometric analysis was performed using a BD FACScan™  Flow Cytometer System.

Acetone-fixed, frozen tissue section of human lymphoma stained with Purified Mouse Anti-Human Perforin  (Cat. No. 556434) using a DAB chromogen and Hematoxylin counterstain. Staining is seen in scattered infiltrated lymphocytes, arrows indicate examples.

Product Details
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BD Pharmingen™
PRF1; P1; PERF; PFN1; PFP; Perforin-1; Cytolysin; FLH2; HPLH2
Human (QC Testing), Cow (Reported)
Mouse BALB/c IgG2b, κ
Purified Granules from the Human Lymphoma Cell Line YT
Intracellular staining (flow cytometry) (Routinely Tested), Immunohistochemistry-frozen, Immunoprecipitation (Tested During Development), Immunofluorescence (Reported), Immunohistochemistry-formalin (antigen retrieval required), Western blot (Not Recommended)
70 kDa
0.5 mg/ml
AB_396418
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.
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Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. 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.
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556434 Rev. 13
Antibody Details
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δG9

Perforin has a key role in cell-mediated cytotoxicity. It is a 70 kDa cytolytic protein that is expressed in the cytoplasmic granules of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. CTLs are involved in eliminating virally infected cells, in anti-tumor immune responses, in allograft rejections, and in some autoimmune diseases. NK cells are important for tumor surveillance and destruction and are involved in allograft rejections. Cytotoxic cells release the contents of their cytotoxic granules, including perforin upon recognition of their target cell. In the presence of calcium, perforin forms transmembrane channels or pores in the membrane of the target cell leading to a cell death that resembles apoptosis. The ability to detect perforin-positive cells with specific antibody should be useful in identifying and understanding perforin-mediated reactions.

Clone δG9 reacts with human and bovine perforin. It does not cross-react with mouse perforin. Purified granules from the human lymphoma cell line YT were used as immunogen. Clone δG9 was initially characterized by immunoprecipitation and immunohistochemistry of frozen tissue sections. The antibody stains scattered lymphocytes in red pulp of spleen, and scattered infiltrated lymphocytes in lymphoma.

556434 Rev. 13
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
556434 Rev.13
Citations & References
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View product citations for antibody "556434" on CiteAb

Development References (6)

  1. Fox WM 3rd, Hameed A, Hutchins GM, et al. Perforin expression localizing cytotoxic lymphocytes in the intimas of coronary arteries with transplant-related accelerated arteriosclerosis. Hum Pathol. 1993; 24(5):477-482. (Clone-specific: Immunohistochemistry). View Reference
  2. Hameed A, Fox WM, Kurman RJ, Hruban RH, Podack ER. Perforin expression in endometrium during the menstrual cycle. Int J Gynecol Pathol. 1995; 14(2):143-150. (Clone-specific: Flow cytometry). View Reference
  3. Hameed A, Fox WM, Kurman RJ, Hruban RH, Podack ER. Perforin expression in human cell-mediated luteolysis. Int J Gynecol Pathol. 1995; 14(2):151-157. (Clone-specific: Immunohistochemistry). View Reference
  4. Hameed A, Olsen KJ, Cheng L, Fox WM 3rd, Hruban RH, Podack ER. Immunohistochemical identification of cytotoxic lymphocytes using human perforin monoclonal antibody. Am J Pathol. 1992; 140(5):1025-1030. (Immunogen: Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
  5. Hameed A, Podack ER, Fox WM, Schafer RW, Sherman ME. Detection of perforin in human peritoneal fluid T-lymphocytes. Acta Cytol. 1996; 40(3):401-407. (Clone-specific: Immunohistochemistry). View Reference
  6. Rukavina D, Balen-Marunic S, Rubesa G, Orlic P, Vujaklija K, Podack ER. Perforin expression in peripheral blood lymphocytes in rejecting and tolerant kidney transplant recipients. Transplantation. 1996; 61(2):285-291. (Clone-specific: Flow cytometry). View Reference
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556434 Rev. 13

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.