RB744 Mouse Anti-Human Perforin

BD Horizon™ RB744 Mouse Anti-Human Perforin

Name: RB744 Mouse Anti-Human Perforin
Brand: RB744 Mouse Anti-Human Perforin
SKU: 570680

Clone δG9 (RUO)

Flow cytometric analysis of Perforin expression in Human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells (PBMC) 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™ RB744 Mouse IgG2b, κ Isotype Control (Cat. No. 570523; dashed line histogram) or BD Horizon™ RB744 Mouse Anti-Human Perforin antibody (Cat. No. 570592/570680; solid line histogram). The fluorescence histogram showing Perforin expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

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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/test

Entrez Gene ID: 5551

RRID: AB_3683687

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

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

RB744 Mouse IgG2b, κ Isotype Control RUO

Size 0.1 mg Cat No. 570523

Fixation and Permeabilization Solution RUO

Size 125 mL Cat No. 554722

Perm/Wash Buffer RUO

Size 100 mL Cat No. 554723

570680 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.

570680 Rev. 1

Format Details

RB744

The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).

Format: RB744

Excitation Source: Blue 488 nm

Excitation Max: 498 nm

Emission Max: 746 nm

570680 Rev.1

Citations & References

View product citations for antibody "570680" on CiteAb

Development References (8)

Baracho GV, Kara N, Rigaud S, Lo E, Widmann SJ, Tyznik AJ. Functional phenotyping of circulating human cytotoxic T cells and NK cells using a 16-color flow cytometry panel.. STAR Protoc. 2022; 3(1):101069. (Clone-specific: Cytotoxicity, Flow cytometry, Functional assay). View Reference
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 (8)

570680 Rev. 1

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