RA (99.9% purity) was purchased from Yuanye Biotechnology, China, and dissolved in dimethyl sulfoxide (DMSO) to a 10mM stock solution that was stored in aliquots in the dark at -20°C. The following antibodies were used for immunoblotting: rabbit anti-actin (Santa Cruz Biotechnology, CA, USA) and anti-PCNA, anti-caspase-3, anti-Bcl-xl, anti-Bcl-2, anti- PARP, anti-STAT3, anti-phospho-STAT3 (Tyr705), anti-JAK2, anti-phospho-JAK2 (Tyr1007/1008), anti- Src, anti-phospho-Src (Tyr416), anti-MDR1, anti-MRP1 (Cell Signaling Technology, Inc., Danvers, MA, USA). Human IL-6 was purchased from Sigma (Sigma-Aldrich, Inc., MO, USA).
Anti mrp1
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-MRP1 is a laboratory reagent used to detect the multidrug resistance-associated protein 1 (MRP1) in biological samples. MRP1 is a membrane transport protein involved in the efflux of various compounds from cells. The Anti-MRP1 product can be used in applications such as Western blotting, immunoprecipitation, and immunohistochemistry to study the expression and localization of MRP1 in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Anti mdr1, by Cell Signaling Technology (2 mentions)
Rabbit anti actin, by Santa Cruz Biotechnology (1 mentions)
Anti src, by Cell Signaling Technology (1 mentions)
Human il 6, by Merck Group (1 mentions)
Anti bcl 2, by Cell Signaling Technology (1 mentions)
Anti phospho jak2, by Cell Signaling Technology (1 mentions)
Anti phospho src, by Cell Signaling Technology (1 mentions)
Anti stat3, by Cell Signaling Technology (1 mentions)
Anti bcl xl, by Cell Signaling Technology (1 mentions)
Anti jak2, by Cell Signaling Technology (1 mentions)
Anti phospho stat3, by Cell Signaling Technology (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Anti parp, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions)
Protein extraction reagent, by Thermo Fisher Scientific (1 mentions)
Infrared scanner, by LI COR (1 mentions)
Anti pgk1, by Santa Cruz Biotechnology (1 mentions)
4 protocols using anti mrp1
1
Molecular Mechanisms of RA in Cancer
2
Quantifying ABC Transporter Protein Expression
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To monitor the expression of ABC transporter proteins, cell monolayers were washed with ice-cold phosphate-buffered saline (PBS) and lysed in Laemmli Sample Buffer (62.5 M Tris-HCl, pH 6.8, 2% SDS, 20% glycerol, 0.2 M dithiothreitol, DTT). Western blot analysis was then performed as previously described [43 (link)] by employing the following antibodies: anti-MDR1 (MDR1/ABCB1 (E1Y7B) rabbit mAb; Cat# 13342) and anti-MRP1 (MRP1/ABCC1 (D7O8N) rabbit mAb; Cat#14685) by Cell Signaling Technology (Euroclone, Pero, MI, Italy; 1:1000), anti-BCRP (ABCG2 (BXP-21); Cat# sc-58222) by Santa Cruz Biotechnology (DBA, Segrate, MI; Italy; 1:500) and α-tubulin (Anti-α-Tubulin, clone B-5-1-2; Cat# T5168) by Sigma-Aldrich (Merck, Milano, MI, Italy; 1:1000), employed as internal loading control. Horseradish peroxidase (HRP)-conjugated secondary antibodies (anti-rabbit and anti-mouse IgG) were provided by Cell Signaling Technology (1:10000). Immunoreactivity was visualized with Immobilon Western Chemiluminescent HRP Substrate (Merck, Milano, MI, Italy).
3
Western Blot Analysis of Drug Resistance Markers
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After incubation, cells were extracted by using protein extraction reagent (Pierce Biotechnology, Rockford, IL, USA). Twenty micrograms of protein was used in SDS—polyacrylamide gel electrophoresis and electrophoretically transferred onto nitrocellulose blots. The nitrocellulose blots were incubated with an anti-MDR1, anti-GSTπ, and anti-MRP1 (Cell Signaling Technology, Inc., Danvers, MA, USA) antibody. The membranes were incubated with horseradish peroxidase-conjugated antibody against mouse, goat, or rabbit IgG (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 1 h. Each membrane was developed with the enhanced chemiluminescence for the detection of the specific antigen.
4
Quantitative Western Blotting Analysis
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The cells were washed twice with PBS,
collected by scraping in radioimmunoprecipitation assay buffer with
EDTA-free complete protease inhibitors (Roche), and sonicated on ice
for 15 s (1 s on/off cycles). Insoluble debris was cleared by centrifugation
at 16,000g and 4 °C for 15 min. The supernatant
was diluted into 4× Laemmli buffer containing 100 mM βME,
heated to 95 °C for 5 min, cooled to room temperature, resolved
on a 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis
gel, and transferred onto a nitrocellulose membrane by standard Western
blotting methods. The membranes were blocked in 2% BSA in TBS + 0.1%
Tween-20 (TBST). The antibodies used in this study include: anti-MRP1
(1:1000, #72202, Cell Signaling Technology) and anti-PGK1 (1:3000,
sc-130335, Santa Cruz Biotechnology). Secondary donkey antirabbit
680 and donkey antimouse 800 (1:10,000, LiCor). The blots were imaged
on a LiCor infrared scanner and images processed in ImageJ.
collected by scraping in radioimmunoprecipitation assay buffer with
EDTA-free complete protease inhibitors (Roche), and sonicated on ice
for 15 s (1 s on/off cycles). Insoluble debris was cleared by centrifugation
at 16,000g and 4 °C for 15 min. The supernatant
was diluted into 4× Laemmli buffer containing 100 mM βME,
heated to 95 °C for 5 min, cooled to room temperature, resolved
on a 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis
gel, and transferred onto a nitrocellulose membrane by standard Western
blotting methods. The membranes were blocked in 2% BSA in TBS + 0.1%
Tween-20 (TBST). The antibodies used in this study include: anti-MRP1
(1:1000, #72202, Cell Signaling Technology) and anti-PGK1 (1:3000,
sc-130335, Santa Cruz Biotechnology). Secondary donkey antirabbit
680 and donkey antimouse 800 (1:10,000, LiCor). The blots were imaged
on a LiCor infrared scanner and images processed in ImageJ.
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