MDR1/ABCB1 Antibody (C219)

Product Discontinued

MDR1/ABCB1 Antibody (C219) Summary

• Immunogen: SDS-solubilized plasma membranes of a multidrug resistant Chinese hamster ovary (CHO) cell line and a human cell line.
• Epitope: This antibody recognizes an internal, highly conserved amino acid sequence: VQEALD and VQAALD, corresponding to the C-terminal and N-terminal regions, respectively, found in both MDR1 and MDR3 isoforms of P Glycoprotein.
• Localization: Cell Membrane; Integral membrane protein
• Specificity: The specific binding of the clone C219 was stronger in epitope sequence VQAALD (N-Terminal fragment) than the epitope sequence VQEALD (C-terminal fragmant) (Georges E., et al., 1990).Analysis of p-glycoprotein expression in skeletal and cardiac muscle revealed that C219 cross reacts with a 200kDa protein that migrates in the same position as myosin. C219 has also been reported to detect the 185kDa c-erbB2 protein (Liu, et al 1997). Care should be taken in the interpretation of true positive P-glycoprotein expression when using myosin-containing tissues or tissues known to have p185 c-erbB2 expression.
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: ABCB1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Block/Neutralize
  • Immunocytochemistry/ Immunofluorescence 1:20
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Paraffin 1:50-1:500
  • Immunoprecipitation 1:200
  • Western Blot 1:200
• Application Notes: IHC-P: Use for Biotin-based detection systems. Sections should be pre-treated with an antigen unmasking solution. *Human liver (positive staining detected along luminal surfaces of bile canaliculi) or human colon (positive staining localized to luminal surface of secretaory epithelium) can be used as positive controls for IHC. **For optimal staining, the primary antibody should be incubated 60 minutes at room temperature. NOTE: Optimal dilutions/concentrations should be determined by the end user. Blocking of ATP hydrolysis by MDR1 confirmed by literature (PMID 1680871, 9016791, 10570132)

Reactivity Notes

Expected to cross-react with wide range of Mammals due to highly conserved epitope sequence. Not yet tested in other species. Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: PBS and BSA
• Preservative: 0.1% Sodium Azide
• Purity: Immunogen affinity purified

Alternate Names for MDR1/ABCB1 Antibody (C219)

• ABC20
• ABCB1
• ABCB1B
• ATP-binding cassette sub-family B member 1
• ATP-binding cassette, sub-family B (MDR/TAP), member 1
• CD243 antigen
• CD243
• CLCS
• colchicin sensitivity
• doxorubicin resistance
• EC 3.6.3
• EC 3.6.3.44
• GP170
• IBD13
• MDR1
• MDR1MGC163296
• multidrug resistance protein 1
• P-glycoprotein 1
• P-gp
• PGY1
• PGY1P-GP

Background

MDR1 is a member of the superfamily of ATP-binding cassette (ABC) transporters. ABC proteins transport various molecules across extra- and intra-cellular membranes. ABC genes are divided into seven distinct subfamilies (ABC1, MDR/TAP, MRP, ALD, OABP, GCN20, White). MDR1 is a member of the MDR/TAP subfamily. Members of the MDR/TAP subfamily are involved in multidrug resistance. The protein encoded by this gene is an ATP-dependent drug efflux pump for xenobiotic compounds with broad substrate specificity. It is responsible for decreased drug accumulation in multidrug-resistant cells and often mediates the development of resistance to anticancer drugs. This protein also functions as a transporter in the blood-brain barrier. (provided by RefSeq)

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for MDR1/ABCB1 Antibody (NB600-1036)(18)

Publications using NB600-1036

• Le Grand M, Kimpton K, Gana C et al. Targeting Functional Activity of AKT Has Efficacy against Aggressive Neuroblastoma ACS Pharmacol Transl Sci 2020-04-08 [PMID: 32259094]
• Tornabene E, Helms HCC, Pedersen SF, Brodin B Effects of oxygen-glucose deprivation (OGD) on barrier properties and mRNA transcript levels of selected marker proteins in brain endothelial cells/astrocyte co-cultures PLoS ONE 2019-08-19 [PMID: 31425564]
• Kao LP, Morad SAF, Davis TS et al. Chemotherapy selection pressure alters sphingolipid composition and mitochondrial bioenergetics in resistant HL-60 cells J. Lipid Res. 2019-09-01 [PMID: 31363040] (WB, Human)
• You, D;Wen, X;Gorczyca, L;Morris, A;Richardson, JR;Aleksunes, LM; Increased MDR1 Transporter Expression in Human Brain Endothelial Cells Through Enhanced Histone Acetylation and Activation of Aryl Hydrocarbon Receptor Signaling Mol. Neurobiol. 2019-04-08 [PMID: 30963442] (WB, Human)
• Bannai T, Mano T, Chen X et al. Chronic cerebral hypoperfusion shifts the equilibrium of amyloid b oligomers to aggregation-prone species with higher molecular weight Sci Rep 2019-02-26 [PMID: 30808940] (WB, Mouse)
• Park S, Majd S. Reconstitution and functional studies of hamster P-glycoprotein in giant liposomes PLoS ONE 2018-06-18 [PMID: 29912971]
• Jandu H, Aluzaite K, Fogh L et al. Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines. BMC Cancer. 2016-01-23 [PMID: 26801902] (WB, Human)
• Wang Z, Xu Y, Meng X et al. Suppression of c-Myc is involved in multi-walled carbon nanotubes' down-regulation of ATP-binding cassette transporters in human colon adenocarcinoma cells. Toxicol. Appl. Pharmacol. 2014-11-22 [PMID: 25461681] (WB, Human)
• Samanta D, Gilkes D, Chaturvedi P, Xiang L, Semenza G. Hypoxia-inducible factors are required for chemotherapy resistance of breast cancer stem cells. Proc Natl Acad Sci U S A. 2014-12-01 [PMID: 25453096] (WB, Human)
• Jeong CB, Kim BM, Kim RK et al. Functional characterization of P-glycoprotein in the intertidal copepod Tigriopus japonicus and its potential role in remediating metal pollution. Aquatic Toxicology. 2014-01-01 [PMID: 25198425]
• Cheung L, Flemming CL, Watt F et al. High-throughput screening identifies Ceefourin 1 and Ceefourin 2 as highly selective inhibitors of multidrug resistance protein 4 (MRP4) Biochem. Pharmacol. 2014-06-25 [PMID: 24973542] (WB, Human)
  
  Details:
  Antibody used in WB in HEK293 (control) and HEK293-Pgp cells (overexpressing P-gp/MDR1) at dilution 1:200 (Figure 3C)
• Halsey CH, Gustafson DL, Rose BJ et al. Development of an in vitro model of acquired resistance to toceranib phosphate (Palladia(R)) in canine mast cell tumor. BMC Vet. Res. 2014-05-06 [PMID: 24885200] (WB, Canine)
• Stein U, Fleuter C, Siegel F et al. Impact of mutant Beta-catenin on ABCB1 expression and therapy response in colon cancer cells Br J Cancer 2012-03-29 [PMID: 22460269] (WB, Human)
• van Den Elsen JM, Kuntz DA, Hoedemaeker FJ et al. Antibody C219 recognizes an alpha-helical epitope on P-glycoprotein. Proc Natl Acad Sci U S A 1999-11-23 [PMID: 10570132]
• Kokubu N, Cohen D, Watanabe T. Functional modulation of ATPase of P-glycoprotein by C219, a monoclonal antibody against P-glycoprotein. Biochem Biophys Res Commun. 1997-01-13 [PMID: 9016791] (B/N)
• Georges E, Zhang JT, Ling V. Modulation of ATP and drug binding by monoclonal antibodies against P-glycoprotein. J Cell Physiol. 1991-09-01 [PMID: 1680871] (B/N)
• Flandrin A. Mise en place d'un nouveau test de permeabilite membranaire a l'aide de la glycoproteine-P reconstituee dans des proteoliposomes Thesis 2017-01-01 (WB, Canine)
• Rutledge ML, Robey-Cafferty SS, Silva EG et al. Monoclonal antibody C219 detection of the multidrug-resistant protein P-glycoprotein in routinely processed tissues: a study of 36 cases of ovarian carcinoma. Mod Pathol;3(3):298-301. 1990-05-01 [PMID: 1972985]

Reviews for MDR1/ABCB1 Antibody (NB600-1036) (2)

Average Rating: 5

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Human, Other.

reviewed by: Mackenzie Malo WB Other 06/13/2016

Summary

• Application: Western Blot
• Sample Tested: OSW canine T-cell lymphoma cells
• Species: Other
• Lot: B201828

reviewed by: Ramanath Narayana Hegde WB Human 05/20/2015

Summary

• Application: Western Blot
• Sample Tested: HeLa Cell Lysate overexpressing MDR1
• Species: Human
• Lot: Lot D11JF02188

FAQs for MDR1/ABCB1 Antibody (NB600-1036). (Showing 1 - 5 of 5 FAQs).

1. I have received MDR1 Antibody NB600-1036; Lot D12DF00689 with no information on the concentration, would you please let me know the concentration of this lot?
   • I am showing that your lot should have a concentration of 72.3 ug/mL.
2. I’m looking at NB600-1036 anti-MDR1 antibody for my work to inhibit PGP ATP binding and hydrolysis. Has this been proven with your company's antibody? I don’t see it being listed as one of the applications. Would you also know how much of the antibody to be added to PGP for inhibition, and also what is the extent of inhibition?
   • NB600-1036 has never been tested for its ability to inhibit ATP binding. Furthermore, the published epitopes recognized by this antibody, VQEALD and VQAALD are not present in the ATP binding domain of MDR1. Please see the Features Section of UniProt P08183. It does not seem that this antibody would be predicted to inhibit ATP binding. However, customer feedback has shown that this clone C219 has inhibited PGP ATP hydrolysis in the following papers: Modulation of ATP and drug binding by monoclonal antibodies against P-glycoprotein. and Functional modulation of ATPase of P-glycoprotein by C219, a monoclonal antibody against P-glycoprotein. As well as a crystal structure about C219 binding to the protein in the following paper: Antibody C219 recognizes an a-helical epitope on P-glycoprotein. Despite the fact that we have never tested this process here at Novus, work published by others would suggest this antibody is capable of inhibition of PGP ATP binding and hydrolysis. Novus encourages feedback on applications that were previously un-tested by our lab through our Innovator's Reward Program.
3. Does this product NB600-1036 react with your purified recombinant MDR1 Protein product H00005243-Q01 ? We want to use the recombinant protein as a positive control for western blot and need to make sure this antibody will detect it.
   • Our product NB600-1036 recognizes two separate epitope regions. One that corresponds to aa 1214-1219 of the human MDR1 protein and one that recognizes aa 571-576 of the human MDR3 protein. These regions are conserved in both proteins. Unfortunately, the partial recombinant protein H00005243-Q01 does not include either of these regions. This protein was made to aa620-710 of the human MDR1 protein. 
4. Before loading the sample, do I need to boil the sample?
   • The protein target of this antibody is MDR1, which is a very large protein with multiple transmembrane domains, according to human proteins. Unfortunately, Novus does not have the standard procedure/recommendation for preparing the lysates. However we have heard many variations of lysate preparations before loading onto the SDS-PAGE gel. For those proteins having lipid moieties linking to the polypeptide chains, boiling but let it sit at room temperature for 30 min is recommended. Since MDR1 is a multi-transmembrane protein, some lipid chains may be attaching to the protein, so you could try this method. We also know that for the big transmembrane proteins, 1% SDS (instead of 0.1% SDS) is used in the RIPA buffer for totally disrupting the protein complexes with the membranes; you could also consider it. Of course the standard method of adding SDS-sample buffer and boiling for 3 min could be tried too. We would recommend using a small amount of antibody for a couple of lysate-SDS preparation methods to optimize the WB experimental conditions.
5. What is the best condition for primary antibody incubation? Is it 4 C overnight or room temperature for 1 hour?
   • We prefer to do the primary antibody treatment at 4 C overnight, due to the fact that the non-specific interaction will be lesser at the lower temperature. However our WB protocol recommends both overnight and 1 hr (at room temperature): Protocol. If you are interested, you could check our paper citations, where this antibody was used in WB: 5 papers; some may be helpful and suggestive. This antibody uses 1:200 dilution.

Bioinformatics

• Gene Symbol: ABCB1
• Entrez:
  • 5243(Human)
  • 24646(Rat)
• Uniprot:
  • P08183(Human)