Invitrogen

Perforin Monoclonal Antibody (eBioOMAK-D), eBioscience™

FIGURE: 1 / 17

Perforin Antibody (14-9392-82) in WB

Immunoblotting of CTLL-2 lysates using Anti-Mouse Perforin Purified at 2 µg/mL and revealed with Anti-Rat HRP.

Perforin Antibody in Flow Cytometry (Flow)

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Product Details

14-9392-82

Applications

Tested Dilution

Publications

Western Blot (WB)

2 µg/mL

View 1 publication 1 publication

Flow Cytometry (Flow)

View 24 publications 24 publications

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host/Isotype

Rat / IgG2a, kappa

Class

Monoclonal

Type

Antibody

Clone

eBioOMAK-D

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

0.5 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_468586

Product Specific Information

Description: The eBioOMAK-D antibody reacts with mouse perforin (pore-forming protein, pfp, Prf). Perforin is one of the cytolytic mediators present in the cytoplasmic granules of cytotoxic T lymphocytes (CTL) and natural killer cells (NK). Perforin is involved in the killing function by CTLs and NKs and has an important role in the immune response against tumors and virus infections.

By immunobloting, eBioOMAK-D recognizes a ~70kDa band in lysates of CTLL-2 mouse cytotoxic cell line and in lysates of IL-2 stimulated but not unstimulated mouse splenocytes. By multi-color intracellular flow cytometric analysis, eBioOMAK-D staining is increased upon stimulation (IL-2 or anti-CD3/28). Intracellular flow staining results showing upregulation of protein expression have been confirmed by immunoblotting. Furthermore, stimulated Perforin Knock-out (developed by Walsh) splenocytes do not stain with eBioOMAK-D nor is any protein detectable by western blotting with eBioOMAK-D as well as other anti-mouse perforin antibodies. Please note that the Kagi perforin knock-out mice may synthesize a truncated form of the protein which may be recognized by eBioOMAK-D.

In IL-2 stimulated mouse splenocytes, NK cells (as determined by CD49b staining) contain perforin while CD8 cells contain little to none and can vary with culture conditions. This has been confirmed by staining and western blotting the two populations using both OMAK-D and P1-8 antibodies. In contrast stimulation of splenocytes with anti-CD3/CD28 antibodies does result in an increase of perforin on both NK cells and CD8 cells.

eBioOMAK-D is also crossreactive to human perforin and co-stains CD56 positive cells in PBMC.

Expression of perforin and Granzyme B do not always correlate (as discussed above in the CD8 population of IL-2 stimulated splenocytes). Granzyme B typically is expressed earlier and at higher levels. Expression of Granzyme B is dramatically increased (more than 10,00 fold based on mRNA estimates and significantly at the protein level based on western blotting and flow analysis) compared to a minimal increase (10-100 fold) in perforin mRNA and protein with IL-2 stimulation.

For intracellular staining and flow cytometric analysis with direct conjugates of anti-mouse perforin, it is highly recommended to use the Foxp3 buffer system (Product # 00-5523). Other buffers may yield varying results.

Applications Reported: This eBioOMAK-D antibody has been reported for use in immunoblotting (WB).

Applications Tested: This eBioOMAK-D antibody has been tested by immunblotting of CTLL-2 cell lysates and can be used at less than or equal to 2 µg/mL.

Purity: Greater than 90%, as determined by SDS-PAGE.

Aggregation: Less than 10%, as determined by HPLC.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Perforin is one of the major cytolytic proteins of cytolytic granules. Perforin is a cytolytic mediator and is stored in and released by cytoplasmic granules. Moreover, perforin is involved in immune defense against tumors and virus infections as mediated by cytotoxic lymphocytes. Perforin is a 555 amino acid protein with a 21 amino acid signal peptide, and has a molecular weight of 70 to 75 kD. Perforin is a pore forming protein with a mechanism of transmembrane channel formation similar to C9, and homology between perforin and C9 have been demonstrated. Studies show that perforin is expressed only in killer cell lines and not in helper T lymphocytes or other tumor cells tested. Perforin is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis. Defects in the perforin gene cause familial hemophagocytic lymphohistiocytosis type 2 (HPLH2), a rare and lethal autosomal recessive disorder of early childhood. Alternative splicing results in multiple transcript variants of perforin.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

P. gingivalis Infection Upregulates PD-L1 Expression on Dendritic Cells, Suppresses CD8+ T-cell Responses, and Aggravates Oral Cancer.

Cancer immunology research

Ren J,Han X,Lohner H,Hoyle RG,Li J,Liang S,Wang H

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Wed Mar 01 00:00:00 EST 2023

Combination of oral STING agonist MSA-2 and anti-TGF-β/PD-L1 bispecific antibody YM101: a novel immune cocktail therapy for non-inflamed tumors.

Journal of hematology & oncology

Yi M,Niu M,Wu Y,Ge H,Jiao D,Zhu S,Zhang J,Yan Y,Zhou P,Chu Q,Wu K

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Sat Oct 08 00:00:00 EDT 2022

CD4⁺ CTLs Act as a Key Effector Population for Allograft Rejection of MSCs in a Donor MHC-II Dependent Manner in Injured Liver.

Aging and disease

Shen S,Li Y,Jin M,Fan D,Pan R,Lin A,Chen Y,Xiang L,Zhao RC,Shao J

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Thu Dec 01 00:00:00 EST 2022

DNA Methylation Patterns in T Cells of Naïve and Influenza A Virus-Infected Mice Developmentally Exposed to an Aryl Hydrocarbon Receptor Ligand.

Environmental health perspectives

Burke CG,Myers JR,Post CM,Boulé LA,Lawrence BP

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Fri Jan 01 00:00:00 EST 2021

Aiduqing formula inhibits breast cancer metastasis by suppressing TAM/CXCL1-induced Treg differentiation and infiltration.

Cell communication and signaling : CCS

Li J,Wang S,Wang N,Zheng Y,Yang B,Wang X,Zhang J,Pan B,Wang Z

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Mon Aug 30 00:00:00 EDT 2021

NLRC4 inhibits NLRP3 inflammasome and abrogates effective antifungal CD8⁺ T cell responses.

iScience

Souza COS,Ketelut-Carneiro N,Milanezi CM,Faccioli LH,Gardinassi LG,Silva JS

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Fri Jun 25 00:00:00 EDT 2021

Blocking interaction between SHP2 and PD-1 denotes a novel opportunity for developing PD-1 inhibitors.

EMBO molecular medicine

Fan Z,Tian Y,Chen Z,Liu L,Zhou Q,He J,Coleman J,Dong C,Li N,Huang J,Xu C,Zhang Z,Gao S,Zhou P,Ding K,Chen L

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Mon Jun 08 00:00:00 EDT 2020

Genetic ablation of adipocyte PD-L1 reduces tumor growth but accentuates obesity-associated inflammation.

Journal for immunotherapy of cancer

Wu B,Chiang HC,Sun X,Yuan B,Mitra P,Hu Y,Curiel TJ,Li R

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Sat Aug 01 00:00:00 EDT 2020

Crosstalks between mTORC1 and mTORC2 variagate cytokine signaling to control NK maturation and effector function.

Nature communications

Wang F,Meng M,Mo B,Yang Y,Ji Y,Huang P,Lai W,Pan X,You T,Luo H,Guan X,Deng Y,Yuan S,Chu J,Namaka M,Hughes T,Ye L,Yu J,Li X,Deng Y

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Mon Nov 19 00:00:00 EST 2018

Activated CD8⁺ T cell extracellular vesicles prevent tumour progression by targeting of lesional mesenchymal cells.

Nature communications

Seo N,Shirakura Y,Tahara Y,Momose F,Harada N,Ikeda H,Akiyoshi K,Shiku H

14-9392 was used in Western Blotting to show that CD8+ T cells have the capacity to protect tumour progression by EV-mediated depletion of mesenchymal tumour stromal cells in addition to their conventional direct cytotoxicity against tumour cells.

Tue Jan 30 00:00:00 EST 2018

IL-17RA-Signaling Modulates CD8+ T Cell Survival and Exhaustion During Trypanosoma cruzi Infection.

Frontiers in immunology

Tosello Boari J,Araujo Furlan CL,Fiocca Vernengo F,Rodriguez C,Ramello MC,Amezcua Vesely MC,Gorosito Serrán M,Nuñez NG,Richer W,Piaggio E,Montes CL,Gruppi A,Acosta Rodríguez EV

14-9392 was used in Flow cytometry/Cell sorting to demonstrate that IL-17RA-signalling cytokines sustain pathogen-specific CD8+ T cell immunity.

Thu Oct 03 00:00:00 EDT 2019

Inhibition of the B7-H3 immune checkpoint limits tumor growth by enhancing cytotoxic lymphocyte function.

Cell research

Lee YH,Martin-Orozco N,Zheng P,Li J,Zhang P,Tan H,Park HJ,Jeong M,Chang SH,Kim BS,Xiong W,Zang W,Guo L,Liu Y,Dong ZJ,Overwijk WW,Hwu P,Yi Q,Kwak L,Yang Z,Mak TW,Li W,Radvanyi LG,Ni L,Liu D,Dong C

14-9392-82 was used in Flow Cytometry to indicate that the B7-H3 checkpoint may serve as a novel target for immunotherapy against cancer.

Tue Aug 01 00:00:00 EDT 2017

1810011o10 Rik Inhibits the Antitumor Effect of Intratumoral CD8⁺ T Cells through Suppression of Notch2 Pathway in a Murine Hepatocellular Carcinoma Model.

Frontiers in immunology

Dai K,Huang L,Huang YB,Chen ZB,Yang LH,Jiang YA

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Wed Nov 20 00:00:00 EST 2019

Targeted calcium influx boosts cytotoxic T lymphocyte function in the tumour microenvironment.

Nature communications

Kim KD,Bae S,Capece T,Nedelkovska H,de Rubio RG,Smrcka AV,Jun CD,Jung W,Park B,Kim TI,Kim M

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Mon May 15 00:00:00 EDT 2017

Tristetraprolin inhibits macrophage IL-27-induced activation of antitumour cytotoxic T cell responses.

Nature communications

Wang Q,Ning H,Peng H,Wei L,Hou R,Hoft DF,Liu J

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Wed Oct 11 00:00:00 EDT 2017

NFATc1 controls the cytotoxicity of CD8⁺ T cells.

Nature communications

Klein-Hessling S,Muhammad K,Klein M,Pusch T,Rudolf R,Flöter J,Qureischi M,Beilhack A,Vaeth M,Kummerow C,Backes C,Schoppmeyer R,Hahn U,Hoth M,Bopp T,Berberich-Siebelt F,Patra A,Avots A,Müller N,Schulze A,Serfling E

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Mon Sep 11 00:00:00 EDT 2017

The breast tumor microenvironment alters the phenotype and function of natural killer cells.

Cellular & molecular immunology

Krneta T,Gillgrass A,Chew M,Ashkar AA

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Thu Sep 01 00:00:00 EDT 2016

Tim-3 negatively mediates natural killer cell function in LPS-induced endotoxic shock.

PloS one

Hou H,Liu W,Wu S,Lu Y,Peng J,Zhu Y,Lu Y,Wang F,Sun Z

Published figure using Perforin monoclonal antibody (Product # 14-9392-82) in Flow Cytometry

Mon Jul 06 00:00:00 EDT 2015

High efficiency of antiviral CD4(+) killer T cells.

PloS one

Hildemann SK,Eberlein J,Davenport B,Nguyen TT,Victorino F,Homann D

14-9392 was used in Flow cytometry/Cell sorting to directly compare and contrast the in vivo cytotoxic T lymphocyte function of antiviral CD4+ and CD8+T cell populations, using an established infectious disease model.

Mon Sep 23 00:00:00 EDT 2013

Type-I IFN drives the differentiation of short-lived effector CD8+ T cells in vivo.

European journal of immunology

Wiesel M,Crouse J,Bedenikovic G,Sutherland A,Joller N,Oxenius A

14-9392-82 was used in Flow Cytometry to study in vivo the role of type-I interferon in the differentiation of CD8+ T cells.

Wed Feb 01 00:00:00 EST 2012

In vivo delivery of interferon-α gene enhances tumor immunity and suppresses immunotolerance in reconstituted lymphopenic hosts.

Gene therapy

Narumi K,Udagawa T,Kondoh A,Kobayashi A,Hara H,Ikarashi Y,Ohnami S,Takeshita F,Ochiya T,Okada T,Yamagishi M,Yoshida T,Aoki K

14-9392 was used in Flow cytometry/Cell sorting to examine if a tumour-specific immune response induced by IFN-α can enhance homeostatic proliferation-induced anti-tumour immunity.

Sun Jan 01 00:00:00 EST 2012

The post sepsis-induced expansion and enhanced function of regulatory T cells create an environment to potentiate tumor growth.

Blood

Cavassani KA,Carson WF,Moreira AP,Wen H,Schaller MA,Ishii M,Lindell DM,Dou Y,Lukacs NW,Keshamouni VG,Hogaboam CM,Kunkel SL

14-9392 was used in Flow cytometry/Cell sorting to investigate the contribution of regulatory T cells to post sepsis immune defects and tumour growth.

Thu Jun 03 00:00:00 EDT 2010

Naive tumor-specific CD4(+) T cells differentiated in vivo eradicate established melanoma.

The Journal of experimental medicine

Xie Y,Akpinarli A,Maris C,Hipkiss EL,Lane M,Kwon EK,Muranski P,Restifo NP,Antony PA

14-9392 was used in Flow cytometry/Cell sorting to study the regression of tumours due to naive tumor-specific CD4(+) T cell differentiation.

Mon Mar 15 00:00:00 EDT 2010

Tumor immunity against a simian virus 40 oncoprotein requires CD8+ T lymphocytes in the effector immune phase.

Journal of virology

Lowe DB,Shearer MH,Jumper CA,Bright RK,Kennedy RC

14-9392 was used in Flow cytometry/Cell sorting to identify role of recombinant SV40 Tag protein immunization in initiating a cytotoxic T-lymphocyte response during tumor cell dissemination and growth.

Fri Jan 01 00:00:00 EST 2010

MHC class I Dk locus and Ly49G2+ NK cells confer H-2k resistance to murine cytomegalovirus.

Journal of immunology (Baltimore, Md. : 1950)

Xie X,Stadnisky MD,Brown MG

14-9392 was used in Flow cytometry/Cell sorting to investigate resistance to murine cytomegalovirus, showing that the MHC class I Dk locus and Ly49G2+ NK cells confer H-2k resistance.

Mon Jun 01 00:00:00 EDT 2009

Bioinformatics

Protein Aliases: Cytolysin; Lymphocyte pore-forming protein; OMAK; OTTHUMP00000019759; P1; Perforin-1; Perforin1; PGFL; PIGF; PIGF-2; PLGF; pore forming protein; PRF1 (pore forming protein 1); RP11-710A11.3

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Gene Aliases: Pfn; Pfp; Prf-1; Prf1

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UniProt ID: (Mouse) P10820

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Entrez Gene ID: (Mouse) 18646

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Function(s)

calcium ion binding wide pore channel activity metal ion binding

Process(es)

immunological synapse formation defense response to tumor cell immune response to tumor cell apoptotic process circadian rhythm cytolysis protein homooligomerization defense response to virus transmembrane transport

Performance Guarantee

If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

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Perforin Monoclonal Antibody (eBioOMAK-D), eBioscience™

Perforin Antibody (14-9392-82) in WB

Product Details

14-9392-82

Western Blot (WB)

Flow Cytometry (Flow)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

Shipping conditions

RRID

Product Specific Information

Target Information

Bioinformatics

Performance Guarantee

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