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Prohibitin Recombinant Rabbit Monoclonal Antibody [SR46-02]

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Specification

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Catalog# ET1602-31

Prohibitin Recombinant Rabbit Monoclonal Antibody [SR46-02]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Predicted reactivity

  • Zebrafish

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Prohibitin Recombinant Rabbit Monoclonal Antibody [SR46-02]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Prohibitin aa 147-196 / 272.

Species Reactivity

Human, Mouse, Rat (Predicted: Zebrafish)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IHC-P, IP, FC

Molecular Weight

Predicted band size: 30 kDa

Positive Control

HEK-293 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, Mouse kidney tissue lysate, Rat brain tissue lysate, Rat kidney tissue lysate, hybrid fish (crucian-carp) liver tissue lysates, HepG2, PC-12, NIH/3T3, human liver carcinoma tissue, human kidney tissue, mouse liver tissue, mouse kidney tissue, mouse stomach tissue, mouse heart tissue, human liver tissue.

Conjugation

unconjugated

Clone Number

SR46-02

RRID

AB_3069644

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

  • 1:1,000

  • IF-Cell

  • 1:100-1:1,000

  • IHC-P

  • 1:200-1:1,000

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Prohibitin is an evolutionarily conserved protein that has antiproliferative activity. The gene encoding human prohibitin maps to chromosome 17q21 and is ubiquitously expressed. Prohibitin is a post-synthetically modified protein that is localized in the inner membrane of mitochondria, where it regulates the cell cycle by blocking the transition between the G1 and S phases, and on the plasma membrane of B cells, where it mediates B cell maturation. Prohibitin mRNA and protein levels are high in G1, decline during the S phase, rise again in G2 and decline in M phase, which suggests that prohibitin controls the cell cycle by using both transcriptional and posttranslational mechanisms. Prohibitin is also a potential tumor suppressor protein that binds to retinoblastoma (Rb) and subsequently inhibits the activity of E2F family members in response to specific signaling cascades. Prohibitin 2 is a repressor of estrogen receptor activity, and is required for somatic and germline differentiation in the larval gonad during embryonic development. Mutations in the Prohibitin genes are correlated with breast cancer development and/or progressionin more than 80% of the cell lines analyzed.

Background References

1. Helenius TO et al. Keratin 8 absence down-regulates colonocyte HMGCS2 and modulates colonic ketogenesis and energy metabolism. Mol Biol Cell 26:2298-310 (2015).

2. Neuhaus A et al. A novel Pex14 protein-interacting site of human Pex5 is critical for matrix protein import into peroxisomes. J Biol Chem 289:437-48 (2014).

Sequence Similarity

Belongs to the prohibitin family.

Tissue Specificity

Widely expressed in different tissues.

Subcellular Location

Cell membrane, Cytoplasm, Membrane, Mitochondrion, Mitochondrion inner membrane, Nucleus.

UNIPROT

SwissProt: P35232 Human

SwissProt: P67778 Mouse

SwissProt: P67779 Rat

SwissProt: Q7T1D8 Zebrafish

Synonyms

Epididymis luminal protein 215 antibody

Epididymis secretory sperm binding protein Li 54e antibody

HEL 215 antibody

HEL S 54e antibody

PHB antibody

PHB_HUMAN antibody

PHB1 antibody

Prohibitin antibody

Images

  • Western blot analysis of Prohibitin on different lysates with Rabbit anti-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/1,000 dilution.<br /><br />Lane 1: HEK-293 cell lysate (20 µg/Lane)<br />Lane 2: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 3: PC-12 cell lysate (20 µg/Lane)<br />Lane 4: Mouse kidney tissue lysate (40 µg/Lane)<br />Lane 5: Rat brain tissue lysate (40 µg/Lane)<br />Lane 6: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 30 kDa<br />Observed band size: 30 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><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/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/1,000 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 Prohibitin on different lysates with Rabbit anti-Prohibitin antibody (ET1602-31) at 1/1,000 dilution.

    Lane 1: HEK-293 cell lysate (20 µg/Lane)
    Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 3: PC-12 cell lysate (20 µg/Lane)
    Lane 4: Mouse kidney tissue lysate (40 µg/Lane)
    Lane 5: Rat brain tissue lysate (40 µg/Lane)
    Lane 6: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 30 kDa
    Observed band size: 30 kDa

    Exposure time: 10 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 (ET1602-31) at 1/1,000 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.

  • Immunocytochemistry analysis of HepG2 cells labeling Prohibitin with Rabbit anti-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/100 dilution.<br /><br />Cells were fixed in 100% precooled methanol for 5 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-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HepG2 cells labeling Prohibitin with Rabbit anti-Prohibitin antibody (ET1602-31) at 1/100 dilution.

    Cells were fixed in 100% precooled methanol for 5 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-Prohibitin antibody (ET1602-31) 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.

  • Immunocytochemistry analysis of PC-12 cells labeling Prohibitin with Rabbit anti-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/1,000 dilution.<br /><br />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-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling Prohibitin with Rabbit anti-Prohibitin antibody (ET1602-31) at 1/1,000 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-Prohibitin antibody (ET1602-31) at 1/1,000 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 (HA601187, 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 liver carcinoma tissue with Rabbit anti-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/200 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/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/200 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 liver carcinoma tissue with Rabbit anti-Prohibitin antibody (ET1602-31) at 1/200 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 (ET1602-31) at 1/200 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 kidney tissue with Rabbit anti-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/1,000 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/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/1,000 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 kidney tissue with Rabbit anti-Prohibitin antibody (ET1602-31) at 1/1,000 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 (ET1602-31) at 1/1,000 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 liver tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>, 1/400) for 30 minutes 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 liver tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-31, 1/400) for 30 minutes 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 kidney tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>, 1/200) for 30 minutes 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 kidney tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-31, 1/200) for 30 minutes 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 stomach tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>, 1/200) for 30 minutes 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 stomach tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-31, 1/200) for 30 minutes 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 heart tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>, 1/200) for 30 minutes 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 heart tissue using anti-Prohibitin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-31, 1/200) for 30 minutes 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 liver tissue with Rabbit anti-Prohibitin antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/200 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/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>) at 1/200 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 liver tissue with Rabbit anti-Prohibitin antibody (ET1602-31) at 1/200 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 (ET1602-31) at 1/200 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 HepG2 cells labeling Prohibitin.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a>, 1μg/mL) (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 HepG2 cells labeling Prohibitin.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1602-31, 1μg/mL) (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).

  • Prohibitin was immunoprecipitated from 0.2 mg HEK-293 cell lysate with <a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HEK-293 cell lysate (input)<br />Lane 2: <a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a> IP in HEK-293 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/ET1602-31" style="font-weight: bold;text-decoration: underline;">ET1602-31</a> in HEK-293 cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 3 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Prohibitin was immunoprecipitated from 0.2 mg HEK-293 cell lysate with ET1602-31 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1602-31 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HEK-293 cell lysate (input)
    Lane 2: ET1602-31 IP in HEK-293 cell lysate
    Lane 3: Rabbit IgG instead of ET1602-31 in HEK-293 cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 3 seconds; ECL: K1801

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Citation