Overview

  • Product name
    Anti-Prohibitin antibody - Mitochondrial Marker
    See all Prohibitin primary antibodies
  • Description
    Rabbit polyclonal to Prohibitin - Mitochondrial Marker
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, Pig
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human Prohibitin.

    Read Abcam's proprietary immunogen policy (Peptide available as ab28492.)

  • Positive control
    • Recombinant Human Prohibitin protein (ab114204) can be used as a positive control in WB. ab28172 antibody gave a positive signal in the following Whole Cell Lysates: HeLa, A431, Jurkat, NIH 3T3, MEF-1 and PC12. In addition, ab28172 antibody gave a positive signal in the following Tissue Lysates: Brain (Mouse), Liver (Mouse), Heart (Mouse), Kidney (Mouse), Pancreas (Mouse), Testis (Mouse), Skeletal Muscle (Mouse), Spinal Cord (Mouse), Ovary (Mouse), Brain (Rat), Liver (Rat) and Heart (Rat).

Properties

Applications

Our Abpromise guarantee covers the use of ab28172 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
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

Images

  • IHC image of Prohibitin staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28172, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate
    Lane 4 : Liver (Mouse) Tissue Lysate - normal tissue
    Lane 5 : Heart (Mouse) Tissue Lysate
    Lane 6 : Kidney (Mouse) Tissue Lysate
    Lane 7 : Mouse pancreas tissue lysate - total protein (ab29363)
    Lane 8 : Testis (Mouse) Tissue Lysate - normal tissue
    Lane 9 : Mouse skeletal muscle tissue lysate - total protein (ab29711)
    Lane 10 : Spinal Cord (Mouse) Tissue Lysate
    Lane 11 : Ovary (Mouse) Tissue Lysate - normal tissue
    Lane 12 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 13 : Brain (Rat) Tissue Lysate - normal tissue
    Lane 14 : Liver (Rat) Tissue Lysate
    Lane 15 : Heart (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

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

  • ICC/IF image of ab28172 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28172, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab28172 staining Prohibitin in mouse liver tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking in 1.5% serum for 10 minutes. The primary antibody was diluted, 1/400 and incubated with sample for 1 hour. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.

    See Abreview

  • All lanes : Anti-Prohibitin antibody - Mitochondrial Marker (ab28172) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : A431 whole cell lysate (ab7909)

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/10000 dilution

    Performed under reducing conditions.

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

  • ICC/IF image of ab28172 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab28172, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used to quench autofluorescence.  5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

References

This product has been referenced in:
  • Festa BP  et al. Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney. Nat Commun 9:161 (2018). Read more (PubMed: 29323117) »
  • Ramseyer VD  et al. Vacuolar protein sorting 13C is a novel lipid droplet protein that inhibits lipolysis in brown adipocytes. Mol Metab 7:57-70 (2018). Read more (PubMed: 29175050) »
See all 39 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for your phone call.

I sincerely apologize for the incorrect advice that was given to you originally.

Out of courtesy and to correct our mistake, I have issued a free of charge replacement with the order number xxx with ab55618, which is better suited for conjugation. You should receive the antibody tomorrow.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

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.

Read More

Answer

Thank you very much for your interest in our antibodies.

We unfortunately do not have small size aliquots of our products, however we can offer you a testing discount on an antibody to test it in a new species. Please see below for further details.

In regards to the choice of prohibitin antibody, I would not recommend to test the ab28172. Indeed, I performed the alignment of the immunogen with the T. brucei sequence, and there is only 50% identity.

I have requested immunogen information for the other antibodies against prohibitin, in order to provide an alignment and to predict crossreactivity. I will get back to you as soon as I have more information. Many of our anti prohibitin antibodies are however monoclonal antibodies generated against the whole protein and wedonot know the epitope of these (ab79198, ab1836, ab59773 and ab54330).

If you would liketotest then one of these antibodies, I can offer a discount off a future purchase if you buythat antibodynow, test it inT.bruceiand submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

0.) Wait until I get back to you with some more information on the probable crossreaction of our other antibodies with the T. brucei prohibitin

1. Reply to this e-mail to let me know that you would like to proceed and test the antibody in T. brucei. I will then send a discount code. This code must be issued before purchasingthe antibodyso please wait for my reply before ordering.

2. Purchase the antibody either by phone, fax, or online (www.abcam.com).

3. Test it in T. brucei.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even ifthe antibodyturns out to be unsuitable for T. brucei, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer or in regards to the predicted crossreactivity of the antibodies, and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (C2C12 cell line)
Specification
C2C12 cell line
Fixative
Paraformaldehyde
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C

Abcam user community

Verified customer

Submitted Jan 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (pancreatic)
Specification
pancreatic
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: DAKO Antigen retrieval buffer pH6
Permeabilization
No

Dr. Ilka Werner-Martini

Verified customer

Submitted Oct 15 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (breast cancer)
Specification
breast cancer
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval Solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted May 26 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (liver)
Specification
liver
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 10 minute(s) · Concentration: 1.5%

Abcam user community

Verified customer

Submitted Mar 11 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Oct 03 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up