ABCC1 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 9 and 2 bp deletion in exon 9.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 9 and 2 bp deletion in exon 9
Alternative names
ABC 29, ABCC, ABCC 1, ATP binding cassette transporter variant ABCC1delta ex13, ATP binding cassette transporter variant ABCC1delta ex13&14, ATP binding cassette transporter variant ABCC1delta ex25, ATP binding cassette transporter variant ABCC1delta ex25&26, ATP binding cassette, sub-family C (CFTR/MRP), member 1, ATP-binding cassette sub-family C member 1, DKFZp686N04233, DKFZp781G125, GS X, LTC4 transporter, Leukotriene C(4) transporter, MRP1_HUMAN, Multidrug resistance protein, Multidrug resistance-associated protein 1, Multiple drug resistance associated protein, Multiple drug resistance protein 1
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ABCC1 KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 9 and 2 bp deletion in exon 9.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Immunocytochemistry, Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 9 and 2 bp deletion in exon 9
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- ABCC1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Immunocytochemistry, Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
MRP1 also known as ABCC1 is a transporter protein with a mass of approximately 190 kDa. This protein belongs to the ATP-binding cassette (ABC) transporter family. MRP1 actively exports a variety of substrates from cells by hydrolyzing ATP to ADP and inorganic phosphate. You can find MRP1 expressed in many tissues including the lung testis and kidney. It helps in cellular detoxification by exporting organic anions and other conjugated metabolites.
- Biological function summary
This protein functions as an important determinant of multidrug resistance in cancer treatment. MRP1 can form part of a larger protein complex that includes other transporters like MRP2. Through its activity MRP1 protects tissues from toxic substances by transporting them out of the cell. Its role in transporting glutathione-conjugated compounds highlights its importance in cellular defense mechanisms against oxidative stress and xenobiotics.
- Pathways
MRP1 participates in the glutathione metabolism and the xenobiotic efflux pathways. Both pathways involve cellular detoxification and the accumulation of anomalies can cause harmful effects in the body. MRP1 works with proteins such as GSTP1 which conjugates toxic substances with glutathione preparing them for export by MRP1. This coordination ensures efficient detoxification and protection of cells from damage.
- Associated diseases and disorders
MRP1 has associations with several conditions particularly drug resistance in various cancers and chronic obstructive pulmonary disease (COPD). In cancer MRP1 overexpression often results in reduced treatment efficacy due to chemotherapy drugs being expelled from the cell helping to resist their cytotoxic effects. Research indicates a connection between MRP1 and P-glycoprotein (ABCB1) in cancer drug resistance pointing to a broader resistive mechanism in which multiple transporters are involved.
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6 product images
Flow Cytometry (Intracellular) - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Flow cytometry overlay histogram showing wild-type HeLa (green line) and ABCC1 knockout HeLa cells (ab265256) stained with Anti-MRP1 antibody [EPR22841-78] ab260038 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-MRP1 antibody [EPR22841-78] ab260038) (1x106 in 100μl at 0.2 μg/ml) for 30 min at 22°C.
The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488, pre-adsorbed) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody was Rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody (wild-type HeLa - black line ABCC1 knockout HeLa - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Immunocytochemistry/ Immunofluorescence - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Anti-MRP1 antibody [EPR21062] ab233383 staining MRP1 in wild-type HeLa cells (top panel) and ABCC1 knockout HeLa cells (ab265256) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-MRP1 antibody [EPR21062] ab233383 at 1/100 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
Western blot - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Lanes 1-4: Merged signal (red and green). Green - Anti-MRP1 antibody [EPR21062] ab233383 observed at 250 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.Anti-MRP1 antibody [EPR21062] ab233383 Anti-MRP1 antibody [EPR21062] was shown to specifically react with MRP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265256 (knockout cell lysate Human ABCC1 (MRP1) knockout HeLa cell lysate ab257242) was used. Wild-type and MRP1 knockout samples were subjected to SDS-PAGE. Anti-MRP1 antibody [EPR21062] ab233383 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-MRP1 antibody [EPR21062] (Anti-MRP1 antibody [EPR21062] ab233383) at 1/1000 dilution
Lane 1: Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: MRP1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2: Western blot - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Lane 3: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4: MRP1 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 171 kDa
Observed band size: 250 kDa
Cell Culture - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Representative images of ABCC1 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Allele-1: 14 bp deletion in exon 9.
Sanger Sequencing - Human ABCC1 (MRP1) knockout HeLa cell line (ab265256)
Allele-2: 2 bp deletion in exon 9.
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