Prohibitin 2 Antibody (JE35-46)

Novus Biologicals | Catalog # NBP3-32900

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Human, Mouse, Rat

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # JE35-46 expressed in HEK293
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Product Specifications

Immunogen

Synthetic peptide within Human Prohibitin 2 aa 151-200 / 299. (Uniprot: Q99623)

Localization

Mitochondrion inner membrane, Cytoplasm, Nucleus, Cell membrane.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

33 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Prohibitin 2 Antibody (JE35-46)

Prohibitin 2 Antibody (JE35-46)

Western Blot: Prohibitin 2 Antibody (JE35-46) [NBP3-32900]

Western Blot: Prohibitin 2 Antibody (JE35-46) [NBP3-32900] - Western blot analysis of Prohibitin 2 on different lysates with Rabbit anti-Prohibitin 2 antibody (NBP3-32900) at 1/1,000 dilution.

Lane 1: Huh7 cell lysate (15 ug/Lane)
Lane 2: HeLa cell lysate (15 ug/Lane)
Lane 3: HepG2 cell lysate (15 ug/Lane)
Lane 4: PANC-1 cell lysate (15 ug/Lane)
Lane 5: RAW264.7 cell lysate (15 ug/Lane)
Lane 6: NIH/3T3 cell lysate (15 ug/Lane)
Lane 7: PC-12 cell lysate (15 ug/Lane)
Lane 8: Mouse liver tissue lysate (30 ug/Lane)
Lane 9: Rat liver tissue lysate (30 ug/Lane)
Lane 10: Mouse heart tissue lysate (30 ug/Lane)
Lane 11: Rat heart tissue lysate (30 ug/Lane)
Lane 12: Mouse testis tissue lysate (30 ug/Lane)
Lane 13: Rat testis tissue lysate (30 ug/Lane)

Predicted band size: 33 kDa
Observed band size: 33 kDa

Exposure time: 3 minutes;

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 (NBP3-32900) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
Prohibitin 2 Antibody (JE35-46)

Immunohistochemistry: Prohibitin 2 Antibody (JE35-46) [NBP3-32900]

Immunohistochemistry: Prohibitin 2 Antibody (JE35-46) [NBP3-32900] - Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Prohibitin 2 antibody (NBP3-32900) 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 (NBP3-32900) 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.
Prohibitin 2 Antibody (JE35-46)

Immunocytochemistry/ Immunofluorescence: Prohibitin 2 Antibody (JE35-46) [NBP3-32900]

Immunocytochemistry/ Immunofluorescence: Prohibitin 2 Antibody (JE35-46) [NBP3-32900] - Immunocytochemistry analysis of HeLa cells labeling Prohibitin 2 with Rabbit anti-Prohibitin 2 antibody (NBP3-32900) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Prohibitin 2 antibody (NBP3-32900) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
Prohibitin 2 Antibody (JE35-46)

Flow Cytometry: Prohibitin 2 Antibody (JE35-46) [NBP3-32900]

Flow Cytometry: Prohibitin 2 Antibody (JE35-46) [NBP3-32900] - Flow cytometric analysis of HeLa cells labeling Prohibitin 2.

Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32900, 1ug/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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Applications for Prohibitin 2 Antibody (JE35-46)

Application
Recommended Usage

Flow Cytometry

1:500-1:1000

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry-Paraffin

1:200-1:1000

Western Blot

1:1000

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Advanced Features

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Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

1*TBS (pH7.4), 0.05% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Prohibitin 2

REA, or repressor of estrogen receptor activity, (also known as BAP37, or B-cell receptor-associated protein) is a 37 kD erythrocyte band 7 integral membrane protein that contains an SPFH domain. It is located in the inner mitochondrial membrane that selectively potentiates the inhibitory effectiveness of antiestrogen-occupied ER. REA plays a possible role in cell aging and acts as a mitochondrial protein chaperone. This protein has been shown to interact with the estrogen receptor, prohibitin, and IgM.

Alternate Names

BAP, Bap37, BCAP37, D-Prohibitin, p22, PHB2, PNAS-141, REA

Gene Symbol

PHB2

Additional Prohibitin 2 Products

Product Documents for Prohibitin 2 Antibody (JE35-46)

Certificate of Analysis

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Product Specific Notices for Prohibitin 2 Antibody (JE35-46)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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