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Ab3368

Manufactured by Abcam
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Sourced in United Kingdom

Ab3368 is an antibody product manufactured by Abcam. It is intended for use in research applications.

Automatically generated - may contain errors

Lab products found in correlation

Hrp substrate, by Merck Group (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Sc 722, by Santa Cruz Biotechnology (1 mentions) Ab2302, by Abcam (1 mentions) Luminata forte western hrp substrate, by Merck Group (1 mentions) Gapdh g8795, by Merck Group (1 mentions) Ab133602, by Abcam (1 mentions) Ab108985, by Abcam (1 mentions) Ab81289, by Abcam (1 mentions) Bxp 53, by Enzo Life Sciences (1 mentions) Ab8227, by Abcam (1 mentions) Bcl 2, by Abcam (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions)

3 protocols using ab3368

1

Quantification and Analysis of Protein Markers

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Proteins were extracted from cell pellets lysed and quantified using the Pierce BCA Protein Assay kit (Thermo Scientific, Rockford, IL, EUA). After that, proteins were separated by electrophoresis on an SDS-polyacrylamide gel and transferred to a nitrocellulose membrane (GE Healthcare, Waukesha, WI, USA). Membranes were blocked for 1 h in 5% (w/v) milk powder in PBS and incubated overnight at 4 °C with primary antibody against anti-NRF2 (1:500) (#sc-722 Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-MRP1 (1:1000) (#ab3368 Abcam, Cambridge, UK), anti-caspase 3 cleaved (1:200) (#ab2302 Abcam), and anti-βactin (1:1000) (#4967 Cell Signaling). Then, membranes were incubated with the correspondent secondary antibody and a chemiluminescent HRP substrate (Merck Millipore. Burlington, MA, USA) was used to develop the membranes. Each blot was performed twice. The full and uncropped western blots can be found in Supplemental Material.
de Souza I., Monteiro L.K., Guedes C.B., Silva M.M., Andrade-Tomaz M., Contieri B., Latancia M.T., Mendes D., Porchia B.F., Lazarini M., Gomes L.R, & Rocha C.R. (2022). High levels of NRF2 sensitize temozolomide-resistant glioblastoma cells to ferroptosis via ABCC1/MRP1 upregulation. Cell Death & Disease, 13(7), 591.
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2

Western Blot Protocol for Protein Detection

Cited 1 time
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Unless otherwise stated, all Western blotting was performed using equipment from BioRad as previously described [30 (link)]. Primary antibodies were used to detect BCRP (BXP-53, 1:5000; Enzo Life Sciences, Farmingdale, NY), β-actin (ab8227, 1:2000, Abcam, Cambridge, MA), transferrin receptor 1 (TFR-1, ab108985, 1:5000 Abcam), placental alkaline phosphatase (PLAP, ab133602, 1:10,000, Abcam), multidrug resistance-associated protein 1 (MRP1, ab3368, 1:2000, Abcam), cluster of differentiation 34 (CD34, ab81289 1:10,000, Abcam), histone H2A (25785, 1:1000, Cell Signaling, Danvers, MA), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, G8795, 1:1000, Sigma-Aldrich). HRP-linked secondary antibodies (anti-rabbit, anti-rat or anti-mouse, 1:2000 Sigma-Aldrich) were used to detect primary antibodies. After the addition of a Luminata Forte Western HRP substrate (Millipore, Billerica, MA), chemiluminescent protein-antibody complexes were visualized using a Fluorchem Imager (ProteinSimple, Santa Clara, CA). Semi-quantitative analysis of bands of the blots was performed using AlphaView Software (ProteinSimple).
Szilagyi J.T., Vetrano A.M., Laskin J.D, & Aleksunes L.M. (2017). Localization of the Placental BCRP/ABCG2 Transporter to Lipid Rafts: Role for Cholesterol in Mediating Efflux Activity. Placenta, 55, 29-36.
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3

Quantification of Cellular Protein Expression

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The cellular proteins were extracted and separated electrophoretically via SDS-PAGE, and subsequently probed with antibodies against MRP1 (cat. no. ab3368, 1:1,500; Abcam, Cambridge, UK), P-gp (cat. no. ab129450, 1:3,000; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), mTOR (cat. no. ab2732, 1:1,500, Abcam), Bcl-2 (cat. no. ab59348, 1:1,000; Santa Cruz Biotechnology, Inc.), and anti-GAPDH (cat. no. ab9485, 1:1,000; Santa Cruz Biotechnology, Inc.) as a control. Quantification of the expression levels of these proteins was performed using Quantity-One-Software 29.0 (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Fan H., Zhu J.H, & Yao X.Q. (2018). Knockdown of long non-coding RNA PVT1 reverses multidrug resistance in colorectal cancer cells. Molecular Medicine Reports, 17(6), 8309-8315.
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