Name: Anti-Prohibitin antibody [EP2803Y]
Brand: RabMAb
SKU: ab75766
Rating: 5 (1 reviews)

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Western blot - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP2803Y] to Prohibitin
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
Reacts with: Mouse, Rat, Human

Carrier Free PE

Product name

Anti-Prohibitin antibody [EP2803Y]
See all Prohibitin primary antibodies
Description

Rabbit monoclonal [EP2803Y] to Prohibitin
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: NIH/3T3, MCF7, A431, HEK-293, HEK-293T and Ramos cell lysates; Mouse kidney lysate; Rat brain lysate. IP: HEK-293 whole cell lysate. IHC-P: Human kidney, liver, placenta, breast carinoma and hepatocellular carcinoma tissues; Mouse kidney, stomach and liver tissues; Rat kidney, stomach and liver tissues. ICC/IF: NIH/3T3 cells; HeLa cells. Flow Cyt (intra): HepG2 and A431 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.12% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP2803Y
Isotype

IgG
Research areas

Alternative Versions
- PE Anti-Prohibitin antibody [EP2803Y] (ab210424)
- Anti-Prohibitin antibody [EP2803Y] - BSA and Azide free (ab239865)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- Ramos whole cell lysate (ab3955)
- A-431 whole cell lysate (ab7909)
Recombinant Protein
- Recombinant Human Prohibitin protein (ab140551)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab75766 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Unpurified format: 1/1000
ICC/IF		1/100.
WB		1/10000 - 1/20000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
IP		1/40.
IHC-P		1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Notes
Flow Cyt (Intra) 1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Unpurified format: 1/1000
ICC/IF 1/100.
WB 1/10000 - 1/20000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
IP 1/40.
IHC-P 1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Function

Prohibitin inhibits DNA synthesis. It has a role in regulating proliferation. As yet it is unclear if the protein or the mRNA exhibits this effect. May play a role in regulating mitochondrial respiration activity and in aging.
Tissue specificity

Widely expressed in different tissues.
Sequence similarities

Belongs to the prohibitin family.
Developmental stage

Levels of expression in fibroblasts decrease heterogeneously during cellular aging.
Cellular localization

Mitochondrion inner membrane.
Target information above from: UniProt accession P35232 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 5245 Human
- Entrez Gene: 18673 Mouse
- Entrez Gene: 25344 Rat
- Omim: 176705 Human
- SwissProt: P35232 Human
- SwissProt: P67778 Mouse
- SwissProt: P67779 Rat
- Unigene: 514303 Human
- Unigene: 263862 Mouse
- Unigene: 29754 Rat
see all
Alternative names
- 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
see all

Western blot - Anti-Prohibitin antibody [EP2803Y] (ab75766)

All lanes : Anti-Prohibitin antibody [EP2803Y] (ab75766) at 1/20000 dilution

Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysates
Lane 2 : Mouse kidney tissue lysates
Lane 3 : Rat brain tissue lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 30 kDa
Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

ab75766 staining Prohibitin in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

Negative control 1: PBS in place of primary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Prohibitin antibody [EP2803Y] (ab75766)

ab75766 staining Prohibitin in NIH/3T3 (mouse embryo fibroblast cell line) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 (1/1000) and ab150120 (1/1000) were used as counterstains for primary antibody ab75766 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)
Flow Cytometry (Intracellular) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Overlay histogram showing HepG2 (human liver hepatocellular carcinoma cell line) cells stained with ab75766 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75766, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunoprecipitation - Anti-Prohibitin antibody [EP2803Y] (ab75766)

ab75766 immunoprecipitating Prohibitin. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/10000.

Lane 1: HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate (10ug)
Lane 2: HEK-293 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab75766 in HEK-293 whole cell lysate
Western blot - Anti-Prohibitin antibody [EP2803Y] (ab75766)

All lanes : Anti-Prohibitin antibody [EP2803Y] (ab75766) at 1/20000 dilution

Lane 1 : MCF7 (human breast adenocarcinoma cell line) whole cell lysates
Lane 2 : A431 (human epidermoid carcinoma cell line) whole cell lysates
Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysates

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 30 kDa
Additional bands at: 30 kDa. We are unsure as to the identity of these extra bands.
Western blot - Anti-Prohibitin antibody [EP2803Y] (ab75766)

All lanes : Anti-Prohibitin antibody [EP2803Y] (ab75766) at 1/20000 dilution

Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) cell lysate
Lane 3 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 4 : Ramos (human Burkitt's lymphoma cell line) cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 30 kDa
Observed band size: 30 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

ab75766 staining Prohibitin in mouse kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

Negative control 1: PBS in place of primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

ab75766 staining Prohibitin in human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody.

Negative control 1: PBS in place of primary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse stomach tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal liver tissue labelling Prohibitin with ab75766.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal placenta tissue labelling Prohibitin with ab75766.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of breast carcinoma tissue labelling Prohibitin with ab75766.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular tissue labelling Prohibitin with ab75766 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling Prohibitin with ab75766 at 1/100. An Alexa Fluor^® 488-conjugated anti-rabbit IgG was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-Prohibitin antibody [EP2803Y] (ab75766)

Immunocytochemistry/Immunofluorescence analysis of NIH/3T3 (mouse embryo fibroblast cell line) cells labelling Prohibitin with ab75766 at 1/100. An Alexa Fluor^® 488-conjugated anti-rabbit IgG was used as the secondary antibody.
Flow Cytometry (Intracellular) - Anti-Prohibitin antibody [EP2803Y] (ab75766)

ab75766 staining Prohibitin in the human cell line A431 (human epidermoid carcinomacell line) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/120. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/500 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Anti-Prohibitin antibody [EP2803Y] (ab75766)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

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Publishing research using ab75766? Please let us know so that we can cite the reference in this datasheet.

ab75766 has been referenced in 15 publications.

Gómez Rodríguez A et al. Elovl2-Ablation Leads to Mitochondrial Membrane Fatty Acid Remodeling and Reduced Efficiency in Mouse Liver Mitochondria. Nutrients 14:N/A (2022). PubMed: 35276915
Jiang L et al. Proteomic analysis reveals ginsenoside Rb1 attenuates myocardial ischemia/reperfusion injury through inhibiting ROS production from mitochondrial complex I. Theranostics 11:1703-1720 (2021). PubMed: 33408776
Yu H et al. Platelet biomarkers for a descending cognitive function: A proteomic approach. Aging Cell 20:e13358 (2021). PubMed: 33942972
Cheng L et al. Emodin Improves Glucose and Lipid Metabolism Disorders in Obese Mice via Activating Brown Adipose Tissue and Inducing Browning of White Adipose Tissue. Front Endocrinol (Lausanne) 12:618037 (2021). PubMed: 34040579
Ma B et al. The SIAH2-NRF1 axis spatially regulates tumor microenvironment remodeling for tumor progression. Nat Commun 10:1034 (2019). PubMed: 30833558

View all Publications for this product

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Application

Flow Cytometry

Sample

Human Cell (keratinocytes)

Specification

keratinocytes

Fixation

Formaldehyde

Permeabilization

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 13 2011

Question

I would ask wether the ab 75766 recognize prohibitin 1 or 2.

Thank,

Abcam community

Verified customer

Asked on May 03 2017

Answer

I can confirm that ab75766 Prohibitin antibody [EP2803Y] has not been tested for it’s specific cross reactivity to prohibitin 1 and 2 proteins in the laboratories.

I checked the alignment of the immunogen with the human prohibitin 1 and 2 protein sequences:

Immunogen alignment to:

Prohibitin 1 isoform 1 is 100%

Prohibitin 1 isoform 2 is 100%

Prohibitin 2 isoform 1 is 42%

Prohibitin 2 isoform 2 is 42.86%

This indicates that the antibody is specific to Prohibitin 1.

Abcam Scientific Support

Answered on May 03 2017

Question

xx However, as our aim is to use it with T.brucei, we would like to be sure that your product is working with our organism.
xxx

Abcam community

Verified customer

Asked on Feb 13 2012

Answer

Thank you very much for your patience.

I have hear back now from the laboratories in regards to the potential crossreaction of the prohibitin antibodies with T. brucei. Unfortunately none of these antibodies seems to be likely to crossreact with T. brucei. I am very sorry for this. Please find here the detailed comments:

ab115669: theexact sequence of the immunogen is proprietary. Upon alignment however with the T. brucei sequence, the homology is only 60%.
The immunogen of ab70672 it is located within aa 170-220.When comparing this immunogen sequence with the T.brucei sequence, the percentage homology is 47%.
Also for the ab75766, with an immunogen is N terminal, and ab75771, with an immunogen between in the middle, the alignment is not good enough to expect a crossreaction. Please see here the alignment of the human and T. brucei sequence http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20120213-172649-0982-54789458-pg

I am very sorry for not being able to provide you with better news. Please do let us know if there is anything else we could do to help you further.

Abcam Scientific Support

Answered on Feb 13 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Developmental stage

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (15)

Customer reviews and Q&As

Flow Cytometry abreview for Anti-Prohibitin antibody [EP2803Y] - Aminoterminal end