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P-glycoprotein (human) monoclonal antibody (C494)

 
ALX-801-003-C100 1 ml 1,004.00 USD
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Product Specification

Alternative Name:ABCB1, CD243, P-glycoprotein 1
 
Clone:C494
 
Host:Mouse
 
Isotype:IgG2a
 
Immunogen:SDS-solubilized plasma membranes of an MDR (multidrug resistant) Chinese hamster ovary cell line and a human leukemic cell line.
 
UniProt ID:P08183
 
Species reactivity:Human
 
Specificity:Recognizes a gene-specific, internal cellular epitope present only on the human MDR1 isoform of P-glycoprotein.
 
Crossreactivity:Cross-reacts with pyruvate carboxylase (PC), an abundant mitochondrial enzyme. Unequivocal plasma membrane patterns of immunostaining represent true P-glycoprotein expression. Weak homogeneous, cytoplasmic or granular patterns of reactivity may represent staining of the PC cross-reactive epitope rather than positive staining for P-glycoprotein. Does not cross-react with human MDR3 P-glycoprotein.
 
Applications:Flow Cytometry, ICC, IHC (FS), IHC (PS), IP, WB
 
Recommended Dilutions/Conditions:Flow cytometry:
Tissue culture cell lines, blood or bone marrow may be used. Specimens should be fixed in 3.7% formaldehyde for 10 minutes at room temperature to permeabilize the cell membrane. Incubation with approximately 5-10µg of antibody per 1x106 cells for 30-60 minutes at 4°C is suggested. Secondary antibodies (such as FITC-labelled anti-mouse IgG antibodies) available from commercial suppliers may be used for the subsequent steps, but should be titered in order to optimize the particular protocol in use.

Immunohistochemistry (frozen sections, paraffin sections):
Adherent cells grown on coverslips or glass slides, cytospin preparations, frozen sections or paraffin-embedded materials may be used. Permeabilization of the cell membrane is a pre-requisite for exposing the epitope recognized by C494.
Frozen sections of cytospin should be air-dried and fixed in acetone for 10 minutes at -20°C.
Paraffin sections should be deparaffinized and rehydrated by conventional means. Secondary antibodies and fluorescent or histochemical detection systems available from other suppliers should be titered for optimal staining. Primary antibody may be diluted to >1:20 for biotin based detection systems. For optimal staining the primary antibody shold be incubated 20-60 minutes at room temperature.

Western Blot:
Protein concentration of specimens used should be in the range of 50-150µg per 50µl. There are many commercially available systems which may be used for direct Western blot. The MAb antibody should be used at a concentration of 1-10µl/ml in indirect Western blots. Exact concentration of antibody and incubation time will vary depending on the particular system used.

Optimal conditions must be determined individually for each application.
 
Application Notes:Flow Cytometry: cell permeabilization required.
Immunocytochemistry: Cytospin preparations.
Immunoprecipitation: The C494 MAb can be used to immunoprecipitate P-glycoprotein from lysates of metabolically or surface radiolabelled cells.
 
Positive Control:Cell lines: Appropriate drug-sensitive parental cell lines and their multidrug resistant derivatives prepared by the same methods as the test specimen can be used as control materials.
Tissue: Human liver (positive staining detected along luminal surfaces of bile canaliculi) or human colon (positive staining localized to luminal surface of secretory epithelium) are recommended.
 
Formulation:Liquid. In PBS containing BSA and 0.1% sodium azide.
 
Handling:After opening, prepare aliquots and store at -70°C. Avoid freeze/thaw cycles.
 
Shipping:Shipped on Blue Ice
 
Short Term Storage:+4°C
 
Long Term Storage:-20°C
 

Product Literature References

Polarized location of SLC and ABC drug transporters in monolayer-cultured human hepatocytes: M. Le Vee, et al.; Toxicol. In Vitro 29, 938 (2015), Application(s): Immunofluorescence , Abstract;
P-glycoprotein expression and function in patients with temporal lobe epilepsy: a case-control study: M. Feldmann, et al.; Lancet Neurol. 12, 777 (2013), Application(s): IHC using brain tissue, Abstract;
Up-regulation of drug resistance-related vaults during dendritic cell development: A.B. Schroeijers, et al.; J. Immunol. 168, 1572 (2002), Abstract;
Activation of the human P-glycoprotein ATPase by trypsin: S.L. Nuti, et al.; Biochemistry 39, 3423 (2000), Abstract;
Expression of P-glycoprotein in hepatocellular carcinoma. A determinant of chemotherapy response: I.O. Ng, et al.; Am. J. Clin. Pathol. 113, 355 (2000), Abstract;
Choroid plexus epithelial expression of MDR1 P glycoprotein and multidrug resistance-associated protein contribute to the blood-cerebrospinal-fluid drug-permeability barrier: V.V. Rao, et al.; PNAS 96, 3900 (1999), Abstract; Full Text
Multidrug resistance phenotype in high grade soft tissue sarcoma: correlation of P-glycoprotein immunohistochemistry with pathologic response to chemotherapy: R.E. Jimenez, et al.; Cancer 86, 976 (1999), Abstract;
Multidrug resistance-1 and p-glycoprotein in human chondrosarcoma cell lines: expression correlates with decreased intracellular doxorubicin and in vitro chemoresistance: J.J. Wyman, et al.; J. Orthop. Res. 17, 935 (1999), Abstract;
Detection of P-glycoprotein in frozen and paraffin-embedded gastric adenocarcinoma tissues using a panel of monoclonal antibodies: F.J. Lacueva, et al.; Histopathology 32, 328 (1998), Abstract;
MDR1 gene-specific monoclonal antibody C494 cross-reacts with pyruvate carboxylase: V.V. Rao, et al.; Cancer Res. 54, 1536 (1994), Abstract;
Comparison of an immunoperoxidase "sandwich" staining method and western blot detection of P-glycoprotein in human cell lines and sarcomas: K. Toth, et al.; Am. J. Pathol. 140, 1009 (1992), Abstract;
Detection of P-glycoprotein isoforms by gene-specific monoclonal antibodies: E. Georges, et al.; PNAS 87, 152 (1990), Abstract; Full Text
Detection of P-glycoprotein in multidrug-resistant cell lines by monoclonal antibodies: N. Kartner, et al.; Nature 316, 820 (1985), Abstract;

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