Recombinant 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
Related conjugates and formulations
Overview
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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.
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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.
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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
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.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2803Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
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 |
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Flow Cyt (Intra) |
1/120.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Unpurified format: 1/1000 |
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ICC/IF |
1/100.
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WB |
1/10000 - 1/20000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
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IP |
1/40.
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IHC-P |
1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Notes |
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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. |
Target
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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. - Information by UniProt
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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
see all -
Alternative names
- Epididymis luminal protein 215 antibody
- Epididymis secretory sperm binding protein Li 54e antibody
- HEL 215 antibody
see all
Images
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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.
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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) -
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.
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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 -
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. -
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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (15)
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