Overview

  • Product name

    Anti-P Glycoprotein antibody [F4]
    See all P Glycoprotein primary antibodies
  • Description

    Mouse monoclonal [F4] to P Glycoprotein
  • Host species

    Mouse
  • Specificity

    ab80594 detects soluble P glycoprotein in the cultured media of viable adriamycin-resistant cells, whereas those of the drug-sensitive parent cells contain no detectable level of soluble P glycoprotein. We have data to indicate that this antibody may not cross react with Mouse. However, this has not been conclusively tested and expression levels may vary in certain cell lines/tissues.
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human, Chinese hamster
  • Immunogen

    Human and hamster drug-resistant viable cells followed by crude plasma membranes.

  • Epitope

    Extracellular amino terminal half. This epitope is resistant to formalin fixation and periodate oxidation.
  • Positive control

    • Drug resistant cells. Human kidney or breast carcinoma.

Properties

Applications

Our Abpromise guarantee covers the use of ab80594 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
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

Fixed-cells only. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Energy-dependent efflux pump responsible for decreased drug accumulation in multidrug-resistant cells.
  • Tissue specificity

    Expressed in liver, kidney, small intestine and brain.
  • Involvement in disease

    Genetic variations in ABCB1 are associated with susceptibility to inflammatory bowel disease type 13 (IBD13) [MIM:612244]. Inflammatory bowel disease is characterized by a chronic relapsing intestinal inflammation. It is subdivided into Crohn disease and ulcerative colitis phenotypes. Crohn disease may involve any part of the gastrointestinal tract, but most frequently the terminal ileum and colon. Bowel inflammation is transmural and discontinuous; it may contain granulomas or be associated with intestinal or perianal fistulas. In contrast, in ulcerative colitis, the inflammation is continuous and limited to rectal and colonic mucosal layers; fistulas and granulomas are not observed. Both diseases include extraintestinal inflammation of the skin, eyes, or joints. Crohn disease and ulcerative colitis are commonly classified as autoimmune diseases.
  • Sequence similarities

    Belongs to the ABC transporter superfamily. ABCB family. Multidrug resistance exporter (TC 3.A.1.201) subfamily.
    Contains 2 ABC transmembrane type-1 domains.
    Contains 2 ABC transporter domains.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • ABC20 antibody
    • ABCB1 antibody
    • ATP binding cassette, sub family B (MDR/TAP), member 1 antibody
    • ATP-binding cassette sub-family B member 1 antibody
    • CD243 antibody
    • CLCS antibody
    • Colchicin sensitivity antibody
    • Doxorubicin resistance antibody
    • GP170 antibody
    • MDR1 antibody
    • MDR1_HUMAN antibody
    • Multidrug resistance 1 antibody
    • Multidrug resistance protein 1 antibody
    • P glycoprotein 1 antibody
    • P gp antibody
    • P-glycoprotein 1 antibody
    • PGY1 antibody
    see all

References

This product has been referenced in:

  • Zhang HH  et al. Role of 5-Aza-CdR in mitomycin-C chemosensitivity of T24 bladder cancer cells. Oncol Lett 14:5652-5656 (2017). Read more (PubMed: 29142609) »
  • Mesgari Abbasi M  et al. The Effects of Cetirizine on P-glycoprotein Expression and Function In vitro and In situ. Adv Pharm Bull 6:111-8 (2016). Read more (PubMed: 27123426) »
See all 7 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us.

Below is a link to our thorough western blot guide, which includes tips on sample preparation, running, and transfering large proteins.

https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf

For your specific situation, I recommmend:

1. Using an 8% gel
2. Since your protein is a multi-pass membrane protein, heating at 70C for 5-10 minutes rather than boiling for 5 minutes to avoid aggregation.
3. Including 0.1% SDS in both your running and transfer buffers if you are not already doing so.
4. Performing a wet transfer if you are not already doing so.

Read More

Answer



I can confirm that it should not be necessary to prepare a membrane fraction in order to detect this protein.

In case you are experiencing high background or to enrich the protein when low signal occurs, I would recommend preparing a membrane fraction.

I personally would try a whole cell extract first.

I would like to suggest to decrease the temperature to 65C and to prolong the heating time up to 10 min when preparing the samples. Multi-pass membrane proteins have the unfortunate tendency to aggregate during boiling periods with high temperatures and beta- mercaptoethanol due to their hydrophobic nature.

I hope this information is helpful and wish you good luck with your research.

Read More

Answer

Thank you for passing on that reply.

I am sorry to hear that the customer has still not observed the expected signal with the anti-P Glycoprotein antibody (ab3366).

As I mentioned in my previous email, I would be happy to provide the customer with a different antibody to try. I would suggest that they have a look at the rabbit polyclonal ab129450 or the mouse monoclonal ab80594. Both of these have been used previously and shown to detect the P Glycoprotein in Western blotting.

The use of the mouse monoclonal ab80594 has been described in the following references:

Chu TM et al. Detection of soluble P-glycoprotein in culture media and extracellular fluids. Biochem Biophys Res Commun 203:506-12 (1994). PubMed: 7915520
Chu TM et al. Characterization of a new monoclonal antibody F4 detecting cell surface epitope and P-glycoprotein in drug-resistant human tumor cell lines. Hybridoma 12:417-29 (1993). PubMed: 7503940



Whilst the results observed with the rabbit polyclonal ab129450 can be observed on the datasheet of the antibody:

https://www.abcam.com/ab129450

Although the Western blot for ab129450 is picking up some background, the band of interest is observed strongly and it may well be that with further optimisation of the blocking conditions etc. that the blot would look much cleaner.

Both of these antibodies would again be covered by the Abpromise guarantee were the customer to have any further problems.

I look forward to receiving your reply.

Read More

Answer

Thank you for your enquiry.

I can confirm that the epitope of ab80594 Anti-P Glycoprotein antibody [F4] is within the extracellular amino terminal end of the protein.

I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your enquiry and your interest in our products.

Here are a few antibodies against the following targets which have been tested and characterized for Flow cytometry. Please take a look at the on-line datasheets for further information about tested species/application/purity/concentration.

P-gp (MDR1)



ab3366: https://www.abcam.com/P-Glycoprotein-antibody-JSB-1-ab3366.html

ab3083: https://www.abcam.com/P-Glycoprotein-antibody-265-F4-ab3083.html

ab80594:https://www.abcam.com/P-Glycoprotein-antibody-F4-BSA-and-Azide-free-ab80594.html



MRP1:



ab24102: https://www.abcam.com/MRP1-antibody-MRPm5-ab24102.html



BCRP:



ab24115: https://www.abcam.com/BCRPABCG2-antibody-BXP-53-ab24115.html



If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Answer

Thank you for contacting us. ab3364; anti P Glycoprotein is no longer available in our catalogue. However we have similar products available to buy. The catalogue numbers are ab3083; ab3366; ab10333; ab80594; ab80626. Please check the datasheets of these antibodies and select a product according to your requirements. The anti MDR2 products are ab71792; ab24108 and ab111209 Anti MRP1 products are ab24102; ab63987; ab99531; ab81903 To search different products please type target name in the search box provided on the Abcam homepage at www.abcam.com. After hitting the search button the system will give you the sorted product as per your target request. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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