Human ABCC1 (MRP1) knockout HeLa cell line
- Advanced Validation
- What is this?
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- Flow Cyt (Intra)
Unknown
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 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 (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) (ab150081) was used at 1/2000 for 30 min at 22°C.
Isotype control antibody was Rabbit IgG (monoclonal) (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.
- ICC/IF
Unknown
Immunocytochemistry/ Immunofluorescence - Human ABCC1 (MRP1) knockout HeLa cell line (AB265256)
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 ab233383 at 1/100 dilution and 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) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (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).
- WB
Lab
Western blot - Human ABCC1 (MRP1) knockout HeLa cell line (AB265256)
Lanes 1-4 : Merged signal (red and green). Green - ab233383 observed at 250 kDa. Red - loading control ab7291 observed at 50 kDa.
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 ab257242) was used. Wild-type and MRP1 knockout samples were subjected to SDS-PAGE. ab233383 and 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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/mrp1-antibody-epr21062-ab233383'>ab233383</a>) at 1/1000 dilution
Lane 1:
Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human ABCC1 (MRP1) knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-abcc1-mrp1-knockout-hela-cell-lysate-ab257242'>ab257242</a>) at 20 µg
Lane 3:
Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 4:
Western blot - Human ABCC1 (MRP1) knockout A549 cell lysate (<a href='/en-us/products/cell-lysates/human-abcc1-mrp1-knockout-a549-cell-lysate-ab261680'>ab261680</a>) at 20 µg
Secondary
Lanes 1 - 4:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 4:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 171 kDa
Observed band size: 250 kDa,50 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ABCC1 (MRP1) knockout HeLa cell line (AB265256)
Allele-1 : 14 bp deletion in exon 9.
- Sanger seq
Unknown
Sanger Sequencing - Human ABCC1 (MRP1) knockout HeLa cell line (AB265256)
Allele-2 : 2 bp deletion in exon 9.
- Cell Culture
Unknown
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.
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Advanced Validation
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- 0
![Flow Cytometry (Intracellular) - Anti-MRP1 antibody [EPR22841-78] (AB260038)](https://content.abcam.com/products/images/mrp1-antibody-epr22841-78-ab260038--flow-cytometry-intracellular-img50445.jpg)
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com