Invitrogen

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

16 Published Figures

29 References

View all (50) Perforin antibodies

Datasheet

Protocols

Questions & Answers

Datasheet

Protocols

Questions & Answers

Antibody Testing Data (1)
Published Figures (16)

Perforin Antibody in Flow Cytometry (Flow)

FIGURE: 1 / 17

Perforin Antibody (11-9392-82) in Flow

C57BL/6 splenocytes were cultured with Mouse IL-2 Recombinant Protein (Product # 14-8021-64) for 4 days and surface stained with Anti-Mouse CD49b (Integrin alpha 2) APC (Product # 17-5971-82) and intracellularly stained with 1.0 µg of Anti-Mouse Perforin FITC using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Quadrant lines demarcate Isotype Control staining.

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Product Details

11-9392-82

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1 µg/test

View 28 publications 28 publications

ELISA (ELISA)

View 1 publication 1 publication

Product Specifications

Species Reactivity

Mouse

Published species

Bacteria, Mouse

Host/Isotype

Rat / IgG2a, kappa

Recommended Isotype Control

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

Class

Monoclonal

Type

Antibody

Clone

eBioOMAK-D

Conjugate

FITC FITC FITC

Excitation/Emission Max

498/517 nm View spectra

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, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_465447

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 intracellular staining and 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: 488 nm; Emission: 520 nm; Laser: Blue 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

BRCA1 deficiency in mature CD8⁺ T lymphocytes impairs antitumor immunity.

Journal for immunotherapy of cancer

Wu B,Qi L,Chiang HC,Pan H,Zhang X,Greenbaum A,Stark E,Wang LJ,Chen Y,Haddad BR,Clagett D,Isaacs C,Elledge R,Horvath A,Hu Y,Li R

11-9392-82 was used in Flow cytometry/Cell sorting to report that mouse Brca1 knockout (KO) in mature T lymphocytes compromises host antitumor immune response to transplanted syngeneic mouse mammary tumors.

Wed Feb 01 00:00:00 EST 2023

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 # 11-9392-82) 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 # 11-9392-82) in Flow Cytometry

Sat Oct 08 00:00:00 EDT 2022

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 # 11-9392-82) 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 # 11-9392-82) in Flow Cytometry

Thu Dec 01 00:00:00 EST 2022

Activated Natural Killer Cell Promotes Nonalcoholic Steatohepatitis Through Mediating JAK/STAT Pathway.

Cellular and molecular gastroenterology and hepatology

Wang F,Zhang X,Liu W,Zhou Y,Wei W,Liu D,Wong CC,Sung JJY,Yu J

11-9392-82 was used in Flow cytometry/Cell sorting to aim to investigate the functional significance of NK cells in NASH evolution.

Mon Apr 04 00:00:00 EDT 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

11-9392-82 was used in Flow Cytometry to identify cellular mechanisms that drive impaired CD4+ T-cell responses later in life following maternal exposure to an exogenous aryl hydrocarbon receptor. ligand.

Fri Jan 01 00:00:00 EST 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

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 # 11-9392-82) 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 # 11-9392-82) in Flow Cytometry

Fri Jun 25 00:00:00 EDT 2021

Glutathione Restricts Serine Metabolism to Preserve Regulatory T Cell Function.

Cell metabolism

Kurniawan H,Franchina DG,Guerra L,Bonetti L,-Baguet LS,Grusdat M,Schlicker L,Hunewald O,Dostert C,Merz MP,Binsfeld C,Duncan GS,Farinelle S,Nonnenmacher Y,Haight J,Das Gupta D,Ewen A,Taskesen R,Halder R,Chen Y,Jäger C,Ollert M,Wilmes P,Vasiliou V,Harris IS,Knobbe-Thomsen CB,Turner JD,Mak TW,Lohoff M,Meiser J,Hiller K,Brenner D

11-9392 was used in Flow cytometry/Cell sorting to reveal an unexpected role for GSH in restricting serine availability to preserve Treg functionality.

Tue May 05 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

Published figure using Perforin monoclonal antibody (Product # 11-9392-82) 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

11-9392 was used in Flow cytometry/Cell sorting to uncover a previously unrecognised role of adipocyte PD-L1 in mitigating obesity-related inflammation and metabolic dysfunction.

Sat Aug 01 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

11-9392 was used in Flow cytometry/Cell sorting to uncover a previously unrecognised role of adipocyte PD-L1 in mitigating obesity-related inflammation and metabolic dysfunction.

Sat Aug 01 00:00:00 EDT 2020

Progranulin prevents regulatory NK cell cytotoxicity against antiviral T cells.

JCI insight

Huang A,Shinde PV,Huang J,Senff T,Xu HC,Margotta C,Häussinger D,Willnow TE,Zhang J,Pandyra AA,Timm J,Weggen S,Lang KS,Lang PA

11-9392 was used in Flow cytometry/Cell sorting to show that progranulin (PGRN) can reduce NK cell cytotoxicity through reduction of NK cell expansion, granzyme B transcription, and NK cell-mediated lysis of target cells.

Thu Sep 05 00:00:00 EDT 2019

Co-inhibitory Molecule B7 Superfamily Member 1 Expressed by Tumor-Infiltrating Myeloid Cells Induces Dysfunction of Anti-tumor CD8⁺ T Cells.

Immunity

Li J,Lee Y,Li Y,Jiang Y,Lu H,Zang W,Zhao X,Liu L,Chen Y,Tan H,Yang Z,Zhang MQ,Mak TW,Ni L,Dong C

11-9392 was used in Flow cytometry/Cell sorting to demonstrate that increased B7S1 expression in myeloid cells from human hepatocellular carcinoma is correlated with CD8+ T cell dysfunction.

Tue Apr 17 00:00:00 EDT 2018

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 # 11-9392-82) in Flow Cytometry

Mon Nov 19 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

11-9392 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

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 # 11-9392-82) 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 # 11-9392-82) 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

Published figure using Perforin monoclonal antibody (Product # 11-9392-82) in Flow Cytometry

Mon Sep 11 00:00:00 EDT 2017

Dendritic cells, macrophages, NK and CD8⁺ T lymphocytes play pivotal roles in controlling HSV-1 in the trigeminal ganglia by producing IL1-beta, iNOS and granzyme B.

Virology journal

Lucinda N,Figueiredo MM,Pessoa NL,Santos BS,Lima GK,Freitas AM,Machado AM,Kroon EG,Antonelli LR,Campos MA

11-9392 was used in Flow cytometry/Cell sorting to provide strong evidence that the responses mediated by DCs, Mo/M?, NK and CD8+ T lymphocytes through IL-1β, iNOS and granzyme B production, respectively, together with the production of type I IFN early in the infection, are crucial to host defense against HSV-1.

Tue Feb 21 00:00:00 EST 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

11-9392 was used in Flow cytometry/Cell sorting to investigate the developmental stage of tumour-associated NK cells, showing that the tumour microenvironment alters the phenotype and function of NK cells.

Thu Sep 01 00:00:00 EDT 2016

Amphiregulin promotes the immunosuppressive activity of intrahepatic CD4⁺ regulatory T cells to impair CD8⁺ T-cell immunity against hepatitis B virus infection.

Immunology

Dai K,Huang L,Chen J,Yang L,Gong Z

11-9392 was used in Flow cytometry/Cell sorting to elucidate a novel mechanism contributing to immunotolerance and viral amplification after Hepatitis B virus infection.

Sun Mar 01 00:00:00 EST 2015

Targeting the inhibitory receptor CTLA-4 on T cells increased abscopal effects in murine mesothelioma model.

Oncotarget

Wu L,Wu MO,De la Maza L,Yun Z,Yu J,Zhao Y,Cho J,de Perrot M

11-9392 was used in Flow cytometry/Cell sorting to investigate the local and abscopal effects of targeting the CTLA-4 T cell inhibitory receptor in a murine mesothelioma model.

Wed May 20 00:00:00 EDT 2015

The tyrosine phosphatase PTPN22 discriminates weak self peptides from strong agonist TCR signals.

Nature immunology

Salmond RJ,Brownlie RJ,Morrison VL,Zamoyska R

11-9392 was used in Flow cytometry/Cell sorting to study how loss-of-function PTPN22 alleles can lead to the population expansion of effector and/or memory T cells and a predisposition to human autoimmunity.

Mon Sep 01 00:00:00 EDT 2014

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 # 11-9392-82) in Flow Cytometry

Mon Jul 06 00:00:00 EDT 2015

Liver sinusoidal endothelial cell lectin inhibits CTL-dependent virus clearance in mouse models of viral hepatitis.

Journal of immunology (Baltimore, Md. : 1950)

Liu B,Wang M,Wang X,Zhao D,Liu D,Liu J,Chen PJ,Yang D,He F,Tang L

11-9392 was used in Flow cytometry/Cell sorting to elucidate the role of LSECtin in pathogenesis of viral infection, showing that its absence led to increased frequency in intrahepatic CTLs, leading to accelerated liver adenovirus clearance.

Mon Apr 15 00:00:00 EDT 2013

A Salmonella Typhimurium-Typhi genomic chimera: a model to study Vi polysaccharide capsule function in vivo.

PLoS pathogens

Jansen AM,Hall LJ,Clare S,Goulding D,Holt KE,Grant AJ,Mastroeni P,Dougan G,Kingsley RA

11-9392 was used in Flow cytometry/Cell sorting to demonstrate that human oncostatin M is neuroprotective by recruiting OSMRβ in the setting of ischemic stroke.

Fri Jul 01 00:00:00 EDT 2011

A Salmonella Typhimurium-Typhi genomic chimera: a model to study Vi polysaccharide capsule function in vivo.

PLoS pathogens

Jansen AM,Hall LJ,Clare S,Goulding D,Holt KE,Grant AJ,Mastroeni P,Dougan G,Kingsley RA

11-9392 was used in Flow cytometry/Cell sorting to demonstrate that human oncostatin M is neuroprotective by recruiting OSMRβ in the setting of ischemic stroke.

Fri Jul 01 00:00:00 EDT 2011

Effector/memory but not naive regulatory T cells are responsible for the loss of concomitant tumor immunity.

Journal of immunology (Baltimore, Md. : 1950)

Lin YC,Chang LY,Huang CT,Peng HM,Dutta A,Chen TC,Yeh CT,Lin CY

11-9392 was used in Flow cytometry/Cell sorting to study the contribution of various Treg cell types to the maintenance of concomitant tumour immunity.

Fri May 15 00:00:00 EDT 2009

Impaired NK cytolytic activity and enhanced tumor growth in NK lytic-associated molecule-deficient mice.

Journal of immunology (Baltimore, Md. : 1950)

Hoover RG,Gullickson G,Kornbluth J

11-9392 was used in Flow cytometry/Cell sorting to indicate that NK lytic-associated molecule-deficient mice have diminished capacity to control tumour metastases.

Tue Dec 01 00:00:00 EST 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

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

Perforin Antibody (11-9392-82) in Flow

Product Details

11-9392-82

Flow Cytometry (Flow)

ELISA (ELISA)

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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