PE Perforin Reagent Set — 556437 - BD Biosciences · PE Perforin Reagent Set Product Information...
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BD Pharmingen™ Technical Data Sheet PE Perforin Reagent Set Product Information Material Number: 556437 QC Testing: Human Reactivity: Component: 51-65995X PE Mouse Anti-Human Perforin Description: Size: 100 tests (1 ea) Vol. per Test: 20 µl Clone Name: δG9 Immunogen: Purified Granules from the Human Lymphoma Cell Line YT Isotype: Mouse (BALB/c) IgG2b, κ Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide. Component: 51-66375X PE Mouse IgG2b, κ Isotype Control Description: Size: 100 tests (1 ea) Vol. per Test: 20 µl Clone Name: 27-35 Isotype: Mouse (C.SW) IgG2b, κ Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide. Description 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. Profile of permeabilized peripheral blood lymphocytes analyzed on a BD FACScan™ (BDIS, San Jose, CA). Cells were stained with anti-human perforin-PE (clone δG9; shaded histogram) or with an IgG2b-R-PE isotype control. 556437 Rev. 12 Page 1 of 2
Transcript of PE Perforin Reagent Set — 556437 - BD Biosciences · PE Perforin Reagent Set Product Information...
BD Pharmingen™
Technical Data Sheet
PE Perforin Reagent Set
Product Information
Material Number: 556437
QC Testing: HumanReactivity:
Component: 51-65995X
PE Mouse Anti-Human PerforinDescription:
Size: 100 tests (1 ea)
Vol. per Test: 20 µl
Clone Name: δG9
Immunogen: Purified Granules from the Human Lymphoma Cell Line YT
Isotype: Mouse (BALB/c) IgG2b, κ
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
Component: 51-66375X
PE Mouse IgG2b, κ Isotype ControlDescription:
Size: 100 tests (1 ea)
Vol. per Test: 20 µl
Clone Name: 27-35
Isotype: Mouse (C.SW) IgG2b, κ
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
DescriptionPerforin 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.
Profile of permeabilized peripheral blood lymphocytes
analyzed on a BD FACScan™ (BDIS, San Jose, CA).
Cells were stained with anti-human perforin-PE (clone δG9;
shaded histogram) or with an IgG2b-R-PE isotype control.
556437 Rev. 12 Page 1 of 2
Preparation and StorageStore 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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Application Notes
Application
Intracellular staining (flow cytometry) Routinely Tested
Suggested Companion Products
Catalog Number Name CloneSize
554714 BD Cytofix/Cytoperm™ Fixation/Permeablization Kit 250 tests (none)
554723 Perm/Wash Buffer 100 ml (none)
554722 Fixation and Permeabilization Solution 125 ml (none)
Product NoticesThis reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental
sample (a test).
1.
Source of all serum proteins is from USDA inspected abattoirs located in the United States. 2.
Please refer to www.bdbiosciences.com/pharmingen/protocols 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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at
www.bdbiosciences.com/colors.
5.
An isotype control should be used at the same concentration as the antibody of interest. 6.
ReferencesHameed 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)
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: Immunohistochemistry, Immunoprecipitation)
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)
556437 Rev. 12 Page 2 of 2
