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          Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party scientific content. Neither the content nor the BenchSci technology and processes for selection have been evaluated by us; we are providing them as-is and without warranty of any kind, including for use or application of the Thermo Fisher Scientific products presented.

          • Primary Antibodies ›
          • Perforin Antibodies

          Invitrogen

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

          Advanced Verification
          16 Published Figures
          45 References
          View all (50) Perforin antibodies

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          Datasheet
          Protocols
          Questions & Answers
          Datasheet
          Protocols
          Questions & Answers

          Cite Perforin Monoclonal Antibody (eBioOMAK-D), PE, eBioscience™

          • Antibody Testing Data (1)
          • Published Figures (16)
          • Advanced Verification (1)
          Perforin Antibody in Flow Cytometry (Flow)
          Group 53 Created with Sketch.
          Perforin Antibody in Flow Cytometry (Flow)
          Group 53 Created with Sketch.

          FIGURE: 1 / 18

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          Perforin Antibody (12-9392-82) in Flow

          Intracellular staining of C57BL/6 splenocytes unstimulated (left) or stimulated with Mouse IL-2 Recombinant Protein (Product # 14-8021-64) (right) with Anti-Mouse CD49b (Integrin alpha 2) APC (Product # 17-5971-82) and 0.5 µg of Anti-Mouse Perforin PE using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Quadrant lines demarcate isotype control staining. {{ $ctrl.currentElement.advancedVerification.fullName }} validation info. View more
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          View Product

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          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody in Flow Cytometry (Flow)
          Perforin Antibody

          Product Details

          12-9392-82

          Applications
          Tested Dilution
          Publications

          Flow Cytometry (Flow)

          1 µg/test
          View 45 publications 45 publications
          Product Specifications

          Species Reactivity

          Mouse

          Published species

          Mouse

          Host/Isotype

          Rat / IgG2a, kappa

          Recommended Isotype Control

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

          Class

          Monoclonal

          Type

          Antibody

          Clone

          eBioOMAK-D

          Conjugate

          PE PE PE
          • Unconjugated
          • APC 
          • FITC
          • Request custom conjugation

          Excitation/Emission Max

          565/576 nm View spectra 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_466243

          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 immunoblotting, 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 cross-reactive 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 BestProtocols®: 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: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green 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.

          How to use the Panel Builder

          Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

          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

          View more View less

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

          View more View less

          UniProt ID: (Mouse) P10820

          View more View less

          Entrez Gene ID: (Mouse) 18646

          View more View less

          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
          It has to be done as per old AB suggested Products section.
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          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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