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Cyclophilin F Recombinant Rabbit Monoclonal Antibody [JM71-39]

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Specification

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Catalog# ET1705-7

Cyclophilin F Recombinant Rabbit Monoclonal Antibody [JM71-39]

Application

  • WB

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Cyclophilin F Recombinant Rabbit Monoclonal Antibody [JM71-39]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Cyclophilin F aa 30-79 / 207.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP, FC

Molecular Weight

Predicted band size: 22 kDa

Positive Control

HEK-293 cell lysate, HeLa cell lysate, Mouse liver tissue lysate, Rat liver tissue lysate, human colon carcinoma tissue, human breast carcinoma tissue, mouse brain tissue, human colon tissue, HeLa.

Conjugation

unconjugated

Clone Number

JM71-39

RRID

AB_3070599

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IHC-P

  • 1:50-1:400

  • IP

  • Use at an assay dependent concentration.

  • FC

  • 1:1,000

Target

Function

Cyclophilins are conserved, ubiquitous and abundant cytosolic peptidyl-prolyl cis-trans isomerases that accelerate the isomerization of XaaPro peptide bonds and the refolding of proteins. Human cyclophilin A (CyPA), an intracellular protein of 165 amino acids, is the target of cyclosporin A (CsA) and is encoded by a single unique gene conserved between yeast to humans. Cyclophilin B (CyPB) is secreted in biological fluids such as blood or milk, and binds to a specific receptor present on human peripheral blood lymphocytes and expressed in Jurkat cells, a line of human lymphoblasts. Cyclophilin D (CyP40) is a widely expressed cytoplasmic protein that catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides. Cyclophilin F (CyP3), is also known as PPIF, Cyp-D or Rotamase F, is a 207 amino acid mitochondrial matrix protein involved in the induction of necrotic and apoptotic cell death through the activation of the mitochondrial permeability transition (mPT) pore.

Background References

1. Vial G et al. Liver mitochondrial function in ZDF rats during the early stages of diabetes disease. Physiol Rep 4(3). pii: e12686 (2016).

2. Chourasia AH et al. Mitophagy defects arising from BNip3 loss promote mammary tumor progression to metastasis. EMBO Rep 16:1145-63 (2015).

Sequence Similarity

Belongs to the cyclophilin-type PPIase family.

Post-translational Modification

Deacetylated at Lys-167 by SIRT3.

Subcellular Location

Mitochondrion matrix.

UNIPROT

SwissProt: P30405 Human

SwissProt: Q99KR7 Mouse

SwissProt: P29117 Rat

Synonyms

Cyclophilin 3 antibody

cyclophilin D antibody

Cyclophilin F antibody

Cyp D antibody

CyP M antibody

CyP-D antibody

CyP-M antibody

CYP3 antibody

CypD antibody

hCyP3 antibody

Expand

Images

  • Western blot analysis of Cyclophilin F on different lysates with Rabbit anti-Cyclophilin F antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/2,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: Mouse liver tissue lysate (40 µg/Lane)<br />Lane 4: Rat liver tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 22 kDa<br />Observed band size: 17 kDa<br /><br />Exposure time: 59 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Cyclophilin F on different lysates with Rabbit anti-Cyclophilin F antibody (ET1705-7) at 1/2,000 dilution.

    Lane 1: HEK-293 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: Mouse liver tissue lysate (40 µg/Lane)
    Lane 4: Rat liver tissue lysate (40 µg/Lane)

    Predicted band size: 22 kDa
    Observed band size: 17 kDa

    Exposure time: 59 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1705-7) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Cyclophilin F antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Cyclophilin F antibody (ET1705-7) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-7) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Cyclophilin F antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Cyclophilin F antibody (ET1705-7) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-7) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Cyclophilin F antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/50 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Cyclophilin F antibody (ET1705-7) at 1/50 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-7) at 1/50 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Cyclophilin F antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/400 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Cyclophilin F antibody (ET1705-7) at 1/400 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1705-7) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling Cyclophilin F.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1705-7" style="font-weight: bold;text-decoration: underline;">ET1705-7</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling Cyclophilin F.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-7, 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).

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