P-Glycoprotein Monoclonal Antibody (F4) Catalog Number MA Product data sheet

Transcription

1 Website: thermofisher.com Customer Service (US): ext. 1 Technical Support (US): ext. 441 P-Glycoprotein Monoclonal Antibody (F4) Catalog Number MA Product data sheet Details Size 500 µl Host/Isotope Class Type Clone Immunogen Conjugate Form Concentration Purification Storage buffer Contains Storage Conditions Mouse / IgG1 Monoclonal Antibody F4 Human and hamster drug-resistant viable cells followed by crude plasma membranes Unconjugated Liquid 0.2 mg/ml Protein G PBS, ph 7.4, with 0.2% BSA 0.09% sodium azide 4 C Species Reactivity Species reactivity Hamster, Human Published species Rat, Human, Mouse Tested Applications Dilution * Flow Cytometry (Flow) 2 ug/test Immunocytochemistry (ICC) 1:10-1:100 Immunofluorescence (IF) 1:10-1:100 Western Blot (WB) 1:10-1:200 Published Applications Immunocytochemistry (ICC) See 4 publications below Miscellaneous PubMed (MISC) See 1 publications below Immunohistochemistry (IHC) See 1 publications below Western Blot (WB) See 8 publications below * Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls. Product specific information MA targets p170 in FACS, WB and ICC/IF applications and shows reactivity with Hamster and Human samples. This antibody does not react with mouse tissue in Western blot applications. The MA immunogen is human and hamster drug-resistant viable cells followed by crude plasma membranes. Background/Target Information Multi-drug resistance(mdr) associated p-glycoprotein (p170) causes resistance to various drugs in a number of cancer patients. It exists in a soluble and membrane associated form. Soluble P-glycoprotein is detected in extracellular fluids of cancer patients, such as malignant ascites and serum. The Mr of soluble P-glycoprotein is the same as that of membrane bound P-glycoprotein.

2 Product Images For P-Glycoprotein Monoclonal Antibody (F4) P-Glycoprotein Antibody (MA ) in IF Immunofluorescent analysis of p170 (green) showing staining in the membrane of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X- 100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a p170 monoclonal antibody (Product # MA ) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x. P-Glycoprotein Antibody (MA ) in WB Western blot analysis of p170 was performed by loading 25 ug of HepG2 (lane 1), 293T (lane 2) and mouse liver (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4 C overnight. The membrane was probed with a p170 monoclonal antibody (Product # MA ) at a dilution of 1:100 overnight at 4 C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~170 kda in HepG2 and 293T cells. P-Glycoprotein Antibody (MA ) in Flow Flow cytometry analysis of p170 in Jurkat cells (green) compared to an isotype control (blue). Cells were

3 P-Glycoprotein Antibody (MA ) in Flow Flow cytometry analysis of p170 in Raji cells (green) compared to an isotype control (blue). Cells were P-Glycoprotein Antibody (MA ) in Flow Flow cytometry analysis of p170 in CHO cells (green) compared to an isotype control (blue). Cells were

4 PubMed References For P-Glycoprotein Monoclonal Antibody (F4) 4 Immunocytochemistry References MA was used in immunocytochemistry to examine trafficking of cancer-targeting alkylphosphocholine analogues across the blood brain barrier Human / 1:25 Human / 4 ug/ml Molecular pharmaceutics (Sep 2016; 13: 3341) "Analysis of Cancer-Targeting Alkylphosphocholine Analogue Permeability Characteristics Using a Human Induced Pluripotent Stem Cell Blood-Brain Barrier Model." Author(s):Clark PA,Al-Ahmad AJ,Qian T,Zhang RR,Wilson HK,Weichert JP,Palecek SP,Kuo JS,Shusta EV PubMed Article URL: MA was used in immunocytochemistry and immunohistochemistry to study the expression of the ABCB1 transporter in human fetal neural stem/progenitor cells Journal of neuroscience research (Sep 2009; 87: 2615) "ABCB1 is predominantly expressed in human fetal neural stem/progenitor cells at an early development stage." Author(s):Yamamoto A,Shofuda T,Islam MO,Nakamura Y,Yamasaki M,Okano H,Kanemura Y PubMed Article URL: MA was used in immunocytochemistry to study the effects of the novel anti-psychotic drug perospirane of the expression and pump activity of P-glycoprotein European journal of clinical pharmacology (Jul 2008; 64: 697) "Effect of the novel antipsychotic drug perospirone on P-glycoprotein function and expression in Caco-2 cells." Author(s):Zhou YG,Li KY,Li HD PubMed Article URL: MA was used in immunocytochemistry to study the ability of P-glycoprotein to prevent the transport of brucine through an in vitro blood-brain barrier Rat / Not Cited 1 Miscellaneous PubMed References Human / 1:25 European journal of pharmacology (Sep 2012; 690: 68) "Influence of P-glycoprotein on brucine transport at the in vitro blood-brain barrier." Author(s):Xu DH,Yan M,Li HD,Fang PF,Liu YW PubMed Article URL: 1 Immunohistochemistry References MA was used in immunocytochemistry to describe methods to differentiate and characterize hpsc-derived brain microvascular endothelial cells Methods (San Diego, Calif.) (May 2016; 101: 93) "Differentiation and characterization of human pluripotent stem cell-derived brain microvascular endothelial cells." Author(s):Stebbins MJ,Wilson HK,Canfield SG,Qian T,Palecek SP,Shusta EV PubMed Article URL: MA was used in immunohistochemistry to examine the prognostic significance of multiple genes in osteosarcoma 8 Western Blot References Mouse / Not Cited Acta orthopaedica et traumatologica turcica (May 2009; 43: 28) "[The effect of resistance-related proteins on the prognosis and survival of patients with osteosarcoma: an immunohistochemical analysis]." Author(s):Ozger H,Eralp L,Atalar AC,Toker B,Esberk Ate L,Sungur M,Bilgiç B,Ayan I PubMed Article URL: MA was used in western blot to study differences in the expression of multidrug resistance pumps and MMP activity in endothelial cells from normal brain and glioma Journal of neurochemistry (Jan 2003; 84: 316) "Differences in multidrug resistance phenotype and matrix metalloproteinases activity between endothelial cells from normal brain and glioma." Author(s):Régina A,Demeule M,Bérubé A,Moumdjian R,Berthelet F,Béliveau R

5 MA was used in western blot to study the effect of Hsp90 inhibitors on cervical carcinoma survival and growth in vitro and in vivo Human / 1:1000 Human / 1:500 Rat / 1:500 Rat / Not Cited Rat / 1:1000 Cancer chemotherapy and pharmacology (Apr 2008; 61: 669) "Efficacy of Hsp90 inhibition for induction of apoptosis and inhibition of growth in cervical carcinoma cells in vitro and in vivo." Author(s):Schwock J,Pham NA,Cao MP,Hedley DW PubMed Article URL: MA was used in western blot to investigate the role of HBx protein on the occurrence of multidrug resistance in hepatoma cell line World journal of gastroenterology (Dec 2004; 10: 3522) "Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line." Author(s):Guan J,Chen XP,Zhu H,Luo SF,Cao B,Ding L MA was used in western blot to study the ability of sirna targeted against P-glycoprotein to reverse multi-drug resistance in a human leukemia cell line Cancer gene therapy (Nov 2004; 11: 707) "Reversal of P-glycoprotein-mediated multidrug resistance with small interference RNA (sirna) in leukemia cells." Author(s):Peng Z,Xiao Z,Wang Y,Liu P,Cai Y,Lu S,Feng W,Han ZC PubMed Article URL: MA was used in western blot to study the role of ABCG2 overexpression in the development of irinotecan resistance in colon cancer cells International journal of cancer (May 2004; 109: 848) "ABCG2 overexpression in colon cancer cells resistant to SN38 and in irinotecan-treated metastases." Author(s):Candeil L,Gourdier I,Peyron D,Vezzio N,Copois V,Bibeau F,Orsetti B,Scheffer GL,Ychou M,Khan QA,Pommier Y, Pau B,Martineau P,Del Rio M PubMed Article URL: MA was used in western blot to study the expression and distribution of P-glycoprotein in the rat placenta during pregnancy Reproductive toxicology (Elmsford, N.Y.) (Jan 2005; 18: 785) "P-glycoprotein expression and distribution in the rat placenta during pregnancy." Author(s):Novotna M,Libra A,Kopecky M,Pavek P,Fendrich Z,Semecky V,Staud F PubMed Article URL: MA was used in western blot to study the functional activity of P-glycoprotein in the rat placental barrier The Journal of pharmacology and experimental therapeutics (Jun 2003; 305: 1239) "Examination of the functional activity of P-glycoprotein in the rat placental barrier using rhodamine 123." Author(s):Pavek P,Staud F,Fendrich Z,Sklenarova H,Libra A,Novotna M,Kopecky M,Nobilis M,Semecky V PubMed Article URL: MA was used in western blot to study the effect of dexamethasone on multidrug drug transporter expression in normal rat tissues FEBS letters (Jan 1999; 442: 208) "Dexamethasone modulation of multidrug transporters in normal tissues." Author(s):Demeule M,Jodoin J,Beaulieu E,Brossard M,Béliveau R