Perforin Rabbit Polyclonal Antibody
Catalog# ER1803-77
Perforin Rabbit Polyclonal Antibody
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WB
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IF-Cell
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IHC-P
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FC
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Human
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unconjugated
Overview
Product Name
Perforin Rabbit Polyclonal Antibody
Antibody Type
Rabbit Polyclonal Antibody
Immunogen
Recombinant protein within Human Perforin aa 306-515 / 555.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, FC
Molecular Weight
90 kDa, predicted band size 61 kDa
Positive Control
HepG2 cell lysate, Daudi cell lysate, HDLM-2, human spleen tissue, human esophagus tissue.
Conjugation
unconjugated
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Immunogen affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IF-Cell
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1:100
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IHC-P
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1:50-1:200
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FC
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1:1,000
Target
Function
The protein encoded by this gene has structural and functional similarities to complement component 9 (C9). Like C9, this protein creates transmembrane tubules and is capable of lysing non-specifically a variety of target cells. Plays a key role in secretory granule-dependent cell death, and in defense against virus-infected or neoplastic cells. This protein is one of the main cytolytic proteins of cytolytic granules, and it is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis. Defects in this 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 encoding the same protein. Plays an important role in killing other cells that are recognized as non-self by the immune system, e.g. in transplant rejection or some forms of autoimmune disease. Can insert into the membrane of target cells in its calcium-bound form, oligomerize and form large pores.
Background References
1. Vergelli M et al. Human autoreactive CD4+ T cell clones use perforin- or Fas/Fas ligand-mediated pathways for target cell lysis. J Immunol 158:2756-2761 (1997).
2. Ando K et al. Perforin, Fas/Fas ligand, and TNF-alpha pathways as specific and bystander killing mechanisms of hepatitis C virus-specific human CTL. J Immunol 158:5283-5291 (1997).
Sequence Similarity
Belongs to the complement C6/C7/C8/C9 family.
Post-translational Modification
N-glycosylated.
Subcellular Location
Cell membrane, Endosome, Membrane, Secreted.
UNIPROT
Synonyms
Cytolysin antibody
FLH2 antibody
HPLH2 antibody
Lymphocyte pore-forming protein antibody
P1 antibody
PERF_HUMAN antibody
perforin 1 (pore forming protein) antibody
Perforin 1 antibody
Perforin-1 antibody
PFP antibody
ExpandImages
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Western blot analysis of Perforin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used at a 1:500 dilution in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: HepG2 cell lysate
Lane 2: Daudi cell lysate -
Immunocytochemistry analysis of HDLM-2 cells labeling Perforin with Rabbit anti-Perforin antibody (ER1803-77) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Perforin antibody (ER1803-77) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Perforin antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-77) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human esophagus tissue using anti-Perforin antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (ER1803-77) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of HDLM-2 cells labeling Perforin.
Cells were fixed and permeabilized. Then stained with the primary antibody (ER1803-77, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"