PRODUCT CODE: ET1602-31

Prohibitin Recombinant Rabbit Monoclonal Antibody [SR46-02] (ET1602-31)

  • Zebrafish
  • Recombinant

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

  • Zebrafish

Western blot analysis of Prohibitin 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 (ET1602-31, 1/500) was used 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.<br />
Positive control: <br />
Lane 1: Jurkat cell lysate<br />
Lane 2: mouse kidney tissue lysate<br />
Lane 1: 293 cell lysate<br />
Lane 2: HepG2 cell lysate
  • Western blot analysis of Prohibitin 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 (ET1602-31, 1/500) was used 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.<br />
Positive control: <br />
Lane 1: Jurkat cell lysate<br />
Lane 2: mouse kidney tissue lysate<br />
Lane 1: 293 cell lysate<br />
Lane 2: HepG2 cell lysate
  • Western blot analysis of Prohibitin on hybrid  fish (crucian-carp) liver tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-31, 1/500) was used 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.
  • ICC staining of Prohibitin in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Prohibitin in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-31, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human liver carcinoma 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 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, 1/400) 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 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 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 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.
  • Flow cytometric analysis of Prohibitin was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1602-31, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Western blot analysis of Prohibitin 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 (ET1602-31, 1/500) was used 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: Jurkat cell lysate
Lane 2: mouse kidney tissue lysate
Lane 1: 293 cell lysate
Lane 2: HepG2 cell lysate

Applications

  • WB

  • ICC

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

  • Zebrafish

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Prohibitin Recombinant Rabbit Monoclonal Antibody [SR46-02] (ET1602-31)

Immunogen

Synthetic peptide within human prohibitin aa 147-196 / 272.

Host

Rabbit

Positive Control

Jurkat cell lysate, mouse kidney tissue lysate, 293 cell lysate, HepG2 cell lysate, hybrid fish (crucian-carp) liver tissue lysates, NIH/3T3, HepG2, human liver carcinoma tissue, human kidney tissue, mouse liver tissue, mouse kidney tissue, mouse stomach tissue, mouse heart tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SR46-02

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

30 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:2,000

  • ICC

  • 1:50-1:200

  • IHC-P

  • 1:200-1:500

  • FC

  • 1:50-1:100

  • IP

  • assay-dependent

TARGET

UNIPROT #

PROTEIN NAME

Prohibitin

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

SEQUENCE SIMILARITIES

Belongs to the prohibitin family.

TISSUE SPECIFICITY

Widely expressed in different tissues.

DEVELOPMENTAL STAGE

Levels of expression in fibroblasts decrease heterogeneously during cellular aging.

SUBCELLULAR LOCATION

Mitochondrion inner membrane.

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.