Invitrogen

Perforin Monoclonal Antibody (eBioOMAK-D), APC, eBioscience™

Perforin Antibody in Flow Cytometry (Flow)

FIGURE: 1 / 18

Perforin Antibody (17-9392-80) in Flow

Intracellular staining of C57BL/6 splenocytes cultured with recombinant mouse IL-2 for 4 days with Anti-Mouse CD49b (Integrin alpha 2) PE (Product # 12-5971-82) and 0.5 µg of Anti-Mouse Perforin APC using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Cells in the lymphocyte gate were used for analysis.

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Product Details

17-9392-80

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1 µg/test

View 32 publications 32 publications

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

Rat IgG2a kappa Isotype Control (eBR2a), APC, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

eBioOMAK-D

Conjugate

APC APC APC

Excitation/Emission Max

651/660 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_469514

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 intracellular staining followed by flow cytometric analysis.

Applications Tested: This eBioOMAK-D antibody has been tested by flow cytometric analysis of stimulated mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for intracellular (nuclear) proteins. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Excitation: 633-647 nm; Emission: 660 nm; Laser: Red Laser.

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.

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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

17-9392-80 was used in Flow Cytometry to investigate the effects of Porphyromonas gingivalis infection on PD-L1 expression on dendritic cells and related immune consequences in the infection milieu of oral cancer.

Wed Mar 01 00:00:00 EST 2023

6-Mercaptopurine potently inhibits recruitment of SHP2 by phosphorylated PD-1 to inhibit PD-1 signalling and enhance T cell function.

Immunology

Liu L,Lei Y,Zheng Z,Zhou X,Chen S,Zeng G,Yu L,Wang P,Chen L

17-9392-80 was used in Flow Cytometry to reveal a novel anti-tumour mechanism for the drug 6-mercaptopurine (6-MP) and suggests that 6-MP warrants further clinical evaluation for other tumour types.

Sun Oct 01 00:00:00 EDT 2023

Single-cell analyses reveal cannabidiol rewires tumor microenvironment via inhibiting alternative activation of macrophage and synergizes with anti-PD-1 in colon cancer.

Journal of pharmaceutical analysis

Sun X,Zhou L,Wang Y,Deng G,Cao X,Ke B,Wu X,Gu Y,Cheng H,Xu Q,Du Q,Chen H,Sun Y

17-9392-80 was used in Flow cytometry/Cell sorting to provide new insights into the anti-tumor effects of CBD.

Sat Jul 01 00:00:00 EDT 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 # 17-9392-80) 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 # 17-9392-80) in Flow Cytometry

Thu Dec 01 00:00:00 EST 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 # 17-9392-80) in Flow Cytometry

Thu Dec 01 00:00:00 EST 2022

Human poliovirus receptor contributes to biliary atresia pathogenesis by exacerbating natural-killer-cell-mediated bile duct injury.

Liver international : official journal of the International Association for the Study of the Liver

Li Y,Li TY,Qi Q,Zhang MT,Tong MX,Su PJ,Zhang ZB

17-9392-80 was used in Flow cytometry/Cell sorting to show that poliovirus receptor contributes to BA pathogenesis by regulating NK cell-mediated bile duct injury; PVR has the value as a biomarker of BA.

Thu Dec 01 00:00:00 EST 2022

CD36-mediated ferroptosis dampens intratumoral CD8⁺ T cell effector function and impairs their antitumor ability.

Cell metabolism

Ma X,Xiao L,Liu L,Ye L,Su P,Bi E,Wang Q,Yang M,Qian J,Yi Q

17-9392-80 was used in Flow cytometry/Cell sorting to reveal a new mechanism of CD36 regulating the function of CD8+ effector T cells and therapeutic potential of targeting CD36 or inhibiting ferroptosis to restore T cell function.

Tue May 04 00:00:00 EDT 2021

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 # 17-9392-80) in Flow Cytometry

Fri Jan 01 00:00:00 EST 2021

Fidarestat induces glycolysis of NK cells through decreasing AKR1B10 expression to inhibit hepatocellular carcinoma.

Molecular therapy oncolytics

Wu T,Ke Y,Tang H,Liao C,Li J,Wang L

Published figure using Perforin monoclonal antibody (Product # 17-9392-80) in Flow Cytometry

Fri Dec 17 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 # 17-9392-80) 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 # 17-9392-80) 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

17-9392 was used in Flow cytometry/Cell sorting to determine steps within PD-1 signalling pathway that can be targeted in anticancer drugs.

Mon Jun 08 00:00:00 EDT 2020

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

17-9392 was used in Flow cytometry/Cell sorting to determine steps within PD-1 signalling pathway that can be targeted in anticancer drugs.

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 # 17-9392-80) in Flow Cytometry

Sat Aug 01 00:00:00 EDT 2020

Regulation of antitumour CD8 T-cell immunity and checkpoint blockade immunotherapy by Neuropilin-1.

Nature communications

Leclerc M,Voilin E,Gros G,Corgnac S,de Montpréville V,Validire P,Bismuth G,Mami-Chouaib F

17-9392-80 was used in Flow Cytometry to conclude that Nrp-1 could be a target for developing combined immunotherapies.

Fri Jul 26 00:00:00 EDT 2019

IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity.

Nature

Song M,Sandoval TA,Chae CS,Chopra S,Tan C,Rutkowski MR,Raundhal M,Chaurio RA,Payne KK,Konrad C,Bettigole SE,Shin HR,Crowley MJP,Cerliani JP,Kossenkov AV,Motorykin I,Zhang S,Manfredi G,Zamarin D,Holcomb K,Rodriguez PC,Rabinovich GA,Conejo-Garcia JR,Glimcher LH,Cubillos-Ruiz JR

17-9392-80 was used in Flow Cytometry to investigate how intra-tumoral T cells respond to IRE1α-XBP1 in patients with ovarian cancer.

Mon Oct 01 00:00:00 EDT 2018

A Macrophage Colony-Stimulating-Factor-Producing γδ T Cell Subset Prevents Malarial Parasitemic Recurrence.

Immunity

Mamedov MR,Scholzen A,Nair RV,Cumnock K,Kenkel JA,Oliveira JHM,Trujillo DL,Saligrama N,Zhang Y,Rubelt F,Schneider DS,Chien YH,Sauerwein RW,Davis MM

17-9392-80 was used in Flow Cytometry to point to an M-CSF-producing γδ T cell subset that fulfills a specialized protective role in the later stage of malaria infection when αβ T cells have declined.

Tue Feb 20 00:00:00 EST 2018

Dysfunction of Natural Killer Cells by FBP1-Induced Inhibition of Glycolysis during Lung Cancer Progression.

Cell metabolism

Cong J,Wang X,Zheng X,Wang D,Fu B,Sun R,Tian Z,Wei H

17-9392 was used in Flow cytometry/Cell sorting to identify novel mechanisms in natural killer cell dysfunction.

Tue Aug 07 00:00:00 EDT 2018

The 17,18-epoxyeicosatetraenoic acid-G protein-coupled receptor 40 axis ameliorates contact hypersensitivity by inhibiting neutrophil mobility in mice and cynomolgus macaques.

The Journal of allergy and clinical immunology

Nagatake T,Shiogama Y,Inoue A,Kikuta J,Honda T,Tiwari P,Kishi T,Yanagisawa A,Isobe Y,Matsumoto N,Shimojou M,Morimoto S,Suzuki H,Hirata SI,Steneberg P,Edlund H,Aoki J,Arita M,Kiyono H,Yasutomi Y,Ishii M,Kabashima K,Kunisawa J

17-9392 was used in Flow cytometry/Cell sorting to determine the role of epoxyeicosatetraeonic acid and the underlying mechanisms related to skin inflammation.

Wed Aug 01 00:00:00 EDT 2018

Suppression of autophagy and HCK signaling promotes PTGS2^high FCGR3^- NK cell differentiation triggered by ectopic endometrial stromal cells.

Autophagy

Mei J,Zhou WJ,Zhu XY,Lu H,Wu K,Yang HL,Fu Q,Wei CY,Chang KK,Jin LP,Wang J,Wang YM,Li DJ,Li MQ

17-9392 was used in Flow cytometry/Cell sorting to show that the macroautophagy/autophagy level of endometrial stromal cells from endometriosis decreased under negative regulation of estrogen.

Fri Oct 11 00:00:00 EDT 2019

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 # 17-9392-80) in Flow Cytometry

Mon Nov 19 00:00:00 EST 2018

Long-peptide vaccination with driver gene mutations in p53 and Kras induces cancer mutation-specific effector as well as regulatory T cell responses.

Oncoimmunology

Quandt J,Schlude C,Bartoschek M,Will R,Cid-Arregui A,Schölch S,Reissfelder C,Weitz J,Schneider M,Wiemann S,Momburg F,Beckhove P

17-9392 was used in Flow cytometry/Cell sorting to study the immunotherapeutic potential of a panel of long peptides comprising driver gene mutations in TP35 and KRAS frequently found in gastrointestinal tumours.

Tue Jan 12 00:00:00 EST 2021

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 # 17-9392-80) 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 # 17-9392-80) in Flow Cytometry

Mon May 15 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

Mon Sep 11 00:00:00 EDT 2017

Expression of inhibitory receptors and polyfunctional responses of T cells are linked to the risk of congenital transmission of T. cruzi.

PLoS neglected tropical diseases

Egui A,Lasso P,Thomas MC,Carrilero B,González JM,Cuéllar A,Segovia M,Puerta CJ,López MC

17-9392 was used in Flow cytometry/Cell sorting to suggest that the maternal immune profile of the CD4+ and CD8+ T cells could be a determining factor in the congenital transmission of T. cruzi.

Thu Jun 01 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 # 17-9392-80) in Flow Cytometry

Thu Sep 01 00:00:00 EDT 2016

ILC3 GM-CSF production and mobilisation orchestrate acute intestinal inflammation.

eLife

Pearson C,Thornton EE,McKenzie B,Schaupp AL,Huskens N,Griseri T,West N,Tung S,Seddon BP,Uhlig HH,Powrie F

17-9392 was used in Flow cytometry/Cell sorting to investigate how a small population of innate lymphoid cells (ILCs) has large effects on immune homeostasis, showing that ILC3s produce GM-CSF to orchestrate acute intestinal inflammation.

Mon Jan 18 00:00:00 EST 2016

Amphiregulin activates regulatory T lymphocytes and suppresses CD8+ T cell-mediated anti-tumor response in hepatocellular carcinoma cells.

Oncotarget

Yuan CH,Sun XM,Zhu CL,Liu SP,Wu L,Chen H,Feng MH,Wu K,Wang FB

17-9392 was used in Flow cytometry/Cell sorting to investigate how hepatocellular carcinoma cells alter endogenous anti-tumor immunity and their related signaling pathways.

Tue Oct 13 00:00:00 EDT 2015

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 # 17-9392-80) in Flow Cytometry

Mon Jul 06 00:00:00 EDT 2015

Killer cell lectin-like receptor G1 deficiency significantly enhances survival after Mycobacterium tuberculosis infection.

Infection and immunity

Cyktor JC,Carruthers B,Stromberg P,Flaño E,Pircher H,Turner J

17-9392 was used in Flow cytometry/Cell sorting to investigate the requirement of the T cell differentiation marker KLRG1 during M. tuberculosis infection, showing that deficiency significantly enhances survival.

Mon Apr 01 00:00:00 EDT 2013

Memory type 2 helper T cells induce long-lasting antitumor immunity by activating natural killer cells.

Cancer research

Kitajima M,Ito T,Tumes DJ,Endo Y,Onodera A,Hashimoto K,Motohashi S,Yamashita M,Nishimura T,Ziegler SF,Nakayama T

17-9392 was used in Flow cytometry/Cell sorting to demonstrate that memory Th2 cells can induce antitumor immunity through IL-4-induced activation of NK cells.

Fri Jul 15 00:00:00 EDT 2011

Involvement of CD226+ NK cells in immunopathogenesis of systemic lupus erythematosus.

Journal of immunology (Baltimore, Md. : 1950)

Huang Z,Fu B,Zheng SG,Li X,Sun R,Tian Z,Wei H

17-9392 was used in Flow cytometry/Cell sorting to study the role of CD226+ natural killer cells in the immunopathogenesis of systemic lupus erythematosus.

Tue Mar 15 00:00:00 EDT 2011

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), APC, eBioscience™

Perforin Antibody (17-9392-80) in Flow

Product Details

17-9392-80

Flow Cytometry (Flow)

Species Reactivity

Published species

Host/Isotype

Recommended Isotype Control

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

Shipping conditions

RRID

Product Specific Information

Target Information

How to use the Panel Builder

Bioinformatics

Performance Guarantee

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