Overview

  • Product name
    Anti-P Glycoprotein antibody [4E3]
    See all P Glycoprotein primary antibodies
  • Description
    Mouse monoclonal [4E3] to P Glycoprotein
  • Host species
    Mouse
  • Specificity
    P-glycoprotein - 170 Kda (External Epitope) The 4E3 monoclonal antibody recognizes an epitope of the mdr1 encoded P-glycoprotein located on the exterior surface of the plasma membrane. 4E3 does not cross-react with the mdr3 gene product.
  • Tested applications
    Suitable for: ICC, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Tissue/ cell preparation (Human squamous lung cancer cell line SW-1573/500).

  • Positive control
    • Cell Lines: Drug-sensitive parental cell lines and their multidrug-resistant derivatives. Tissue: Liver (positive staining detected along luminal surfaces of bile canaliculi) or colon (positive staining localized to luminal surface of secretory epithelium).
  • General notes


    P-glycoprotein, the product of the MDR1 gene, is expressed in distinct non-malignant cells, typically cells with secretory and excretory functions. It is assumed to function as an ATP-dependent drug efflux pump with broad substrate specificity. The highest expression of P-glycoprotein has been observed in kidney (proximal tubules), liver (bile canaliculi), adrenal gland and intestine, suggesting that the primary role of P-glycoprotein is in the normal secretion of physiological metabolites and ingested chemicals into bile, urine and the lumen of the intestinal tract. Elevated levels of P-glycoprotein have also been reported in multidrug-resistant cell lines and in colon, endometrial, ovarian, and breast tumors, as well as in sarcomas and leukemias/lymphomas.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity
    Protein G purified
  • Primary antibody notes
    P-glycoprotein, the product of the MDR1 gene, is expressed in distinct non-malignant cells, typically cells with secretory and excretory functions. It is assumed to function as an ATP-dependent drug efflux pump with broad substrate specificity. The highest expression of P-glycoprotein has been observed in kidney (proximal tubules), liver (bile canaliculi), adrenal gland and intestine, suggesting that the primary role of P-glycoprotein is in the normal secretion of physiological metabolites and ingested chemicals into bile, urine and the lumen of the intestinal tract. Elevated levels of P-glycoprotein have also been reported in multidrug-resistant cell lines and in colon, endometrial, ovarian, and breast tumors, as well as in sarcomas and leukemias/lymphomas.
  • Clonality
    Monoclonal
  • Clone number
    4E3
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab10333 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
ICC 1/10.
Flow Cyt 1/10.

ab170191 - Mouse monoclonal IgG2a, 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:
  • Lu WQ  et al. Knockdown of PKM2 and GLS1 expression can significantly reverse oxaliplatin-resistance in colorectal cancer cells. Oncotarget 8:44171-44185 (2017). Read more (PubMed: 28498807) »
  • Cho CF  et al. Blood-brain-barrier spheroids as an in vitro screening platform for brain-penetrating agents. Nat Commun 8:15623 (2017). Human . Read more (PubMed: 28585535) »
See all 9 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Answer

Thank you for your inquiry.


I heard back from the supplying lab that theyhave not mapped the exact epitope for this antibody. We have licensed this from the inventor of the antibody. The only information we have about it was provided by Dr Arceci's lab. The following publication describes the invention of this antibody, however it doesn't have a lot of information regarding epitope.


Arceci RJ, et al. Monoclonal antibody to an external epitope of the human mdr1 P-glycoprotein. Cancer Res. 53:310-317, 1993



I'm sorry I cannot provide more insight into this. I did a quick literature search and it doesn't appear that anyone has published information regarding the epitope of 4E3, so unfortunately we do not know which extracellular epitope it binds. I apologize that we have not been able to help you in this regard, but please let me know if you have any other questions.

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Question
Answer

Thank you for your phone call.
As we discussed, ab10333 has not been validated for use in WB and may or may not be suitable for use in that application. While it will not be covered under our Abpromise guarantee, the following may help to improve your results:
1) Because P Glycoprotein is a multi-pass membrane protein, it will tend to aggregate when boiled. To avoid this, I would recommend heating to a lower temperature (around 70C) for 10-20 minutes to achieve denaturation.
2) P Glycoprotein is also a high molecular weight protein. To run effectively, the percentage of the gel will need to be 8% or lower.
3) High molecular weight proteins may not transfer properly using standard buffers. Adding 0.1% SDS to the transfer buffer and keeping the methanol concentration to 10% or lower will help the transfer efficiency. It is recommended to validate the transfer with a ponceau stain to ensure the larger proteins have transferred properly.
4) In general, you may be able to increase your signal by using more primary antibody or incubating overnight at 4C.
I hope this helps, if not, please provide a brief outline of your protocol information and I will be happy to assist you further.

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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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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (liver, hepatoma, HepG2)
Specification
liver, hepatoma, HepG2
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Tx100
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Aug 31 2007

Answer

Thank you for your email. I do apologize as I think that a reply got lost somehow in the system. For this antibody, we were selling it in a smaller size (100 ul) but it did not provide enough tests. We are now providing it in a 500 ul size. If your customer is interested, I can send you the 500 ul vial free of charge, or I can provide you with a credit. Please let me know which is preferred.

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Answer

Thank you for your enquiry. We have received the following information from the source of ab0333: "We recommend that the customer start with a 1:10 dilution and titer Ab from there for their individual application." In light of this information we can certainly offer you a refund or credit note and we are considering removing the antibody from our catalogue, I am sorry about this inconvenience, Please let me know if your customer wishes a credit note and we will issue it to you,

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Answer

Thank you for your enquiry and for your patience. Unfortunately, none of these antibodies are available in a conjugated form but we do have a wide range of secondary antibodies to choose from (including ones conjugated to FITC and PE). At the bottom of the online datasheets for each of these primary antibodies, there is a list of compatible secondary antibodies that can be used. We unfortunately can't offer any discounts. All the information that we have regarding these antibodies is located on the online datasheets, and ab10333 and ab3373 have been tested for application in FACS but ab3380 and ab3375 have not to our knowledge. If you have any more questions, please contact us again.

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Answer

Thank you for your enquiry regarding ab10333. I recommend you refer to our general flow cytometry protocol for all relevant flow cytometry information. You can view this information by clicking on the protocol tab on our online product datasheet. If you need information other than that listed on our product datasheet, please contact us again and we will gladly assist you.

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

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