BV421 Mouse Anti-Human Perforin

BD Horizon™ BV421 Mouse Anti-Human Perforin

Name: BV421 Mouse Anti-Human Perforin
Brand: BD Horizon™
SKU: 563393

Clone δG9

(RUO)

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 BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; dashed line histogram) or BD Horizon™ BV421 Mouse Anti-Human Perforin antibody (Cat. No. 563393; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

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Product Details

Brand: BD Horizon™

Alternative Name: PRF1; P1; PERF; PFN1; PFP; Perforin-1; Cytolysin; FLH2; HPLH2

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG2b, κ

Immunogen: Purified Granules from the Human Lymphoma Cell Line YT

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 5 µl

RRID: AB_2738178

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: 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 BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

This 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).
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.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Brilliant Violet™ 421 is a trademark of Sirigen.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Fixation and Permeabilization Solution RUO

Size 125 mL Cat No. 554722

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

BV421 Mouse IgG2b, κ Isotype Control RUO

Size 50 µg Cat No. 562748

563393 Rev. 1

Antibody Details

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.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

563393 Rev. 1

Format Details

BV421

The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV421

Excitation Source: Violet 405 nm

Excitation Max: 407 nm

Emission Max: 423 nm

563393 Rev.1

Citations & References

Development References (7)

Endsley JJ, Furrer JL, Endsley MA, et al. Characterization of bovine homologues of granulysin and NK-lysin. J Immunol. 2004; 173(4):2607-2614. (Clone-specific: Flow cytometry). View Reference
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
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
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
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). View Reference
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
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

View All (7)

563393 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.