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AB261855

Human ABCC2 knockout A549 cell line

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ABCC2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 11 bp deletion Frameshift = 97%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
6 Images
Western blot - Human ABCC2 knockout A549 cell line (AB261855)
  • WB

Lab

Western blot - Human ABCC2 knockout A549 cell line (AB261855)

Lanes 1 - 4 : Merged signal (red and green). Green - ab172630 observed at 210 kDa. Red - loading control ab130007 observed at 130 kDa.

ab172630 was shown to recognize MRP2 in wild-type A549 cells as signal was lost at the expected MW in abCC2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and abCC2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab172630 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4° at 1/1000 dilution and 1/1000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MRP2 antibody [EPR10998] (<a href='/en-us/products/primary-antibodies/mrp2-antibody-epr10998-ab172630'>ab172630</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

ABCC2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ABCC2 knockout A549 cell line (ab261855)

Lane 3:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 174 kDa

false

Western blot - Human ABCC2 knockout A549 cell line (AB261855)
  • WB

Lab

Western blot - Human ABCC2 knockout A549 cell line (AB261855)

Lanes 1 - 3 : Merged signal (red and green). Green - ab3373 observed at 210 kDa. Red - loading control ab52866 observed at 50 kDa.

ab3373 was shown to recognize abCC2 in wild-type A549 cells as signal was lost at the expected MW in abCC2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and abCC2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab3373 and ab52866 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4° at 1/20 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MRP2 antibody [M2 III-6] (<a href='/en-us/products/primary-antibodies/mrp2-antibody-m2-iii-6-ab3373'>ab3373</a>) at 1/20 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

ABCC2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ABCC2 knockout A549 cell line (ab261855)

Lane 3:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 174 kDa

false

Western blot - Human ABCC2 knockout A549 cell line (AB261855)
  • WB

Lab

Western blot - Human ABCC2 knockout A549 cell line (AB261855)

Lanes 1 - 4 : Merged signal (red and green). Green - ab187644 observed at 210 kDa. Red - loading control ab130007 observed at 130 kDa.

ab187644 was shown to recognize MRP2 in wild-type A549 cells as signal was lost at the expected MW in abCC2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and abCC2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab187644 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4° at 1/1000 dilution and 1/1000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MRP2 antibody [EPR10997(2)] (<a href='/en-us/products/primary-antibodies/mrp2-antibody-epr109972-ab187644'>ab187644</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

ABCC2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ABCC2 knockout A549 cell line (ab261855)

Lane 3:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 174 kDa

false

Western blot - Human ABCC2 knockout A549 cell line (AB261855)
  • WB

Lab

Western blot - Human ABCC2 knockout A549 cell line (AB261855)

Lanes 1 - 2 : Merged signal (red and green). Green - ab15603 observed at 210 kDa. Red - loading control ab52866 observed at 50 kDa.

ab15603 was shown to recognize abCC2 in wild-type A549 cells as signal was lost at the expected MW in abCC2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and abCC2 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab15603 and ab52866 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4° at 1/20 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MRP2 antibody [M2III-5] (<a href='/en-us/products/primary-antibodies/mrp2-antibody-m2iii-5-ab15603'>ab15603</a>) at 1/20 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

ABCC2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ABCC2 knockout A549 cell line (ab261855)

Predicted band size: 174 kDa

false

Next Generation Sequencing - Human ABCC2 knockout A549 cell line (AB261855)
  • NGS

Supplier Data

Next Generation Sequencing - Human ABCC2 knockout A549 cell line (AB261855)

11 bp deletion after Phe130 of the WT protein

Next Generation Sequencing - Human ABCC2 knockout A549 cell line (AB261855)
  • NGS

Lab

Next Generation Sequencing - Human ABCC2 knockout A549 cell line (AB261855)

X = 11 bp deletion

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 11 bp deletion Frameshift = 97%

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB172630

Anti-MRP2 antibody [EPR10998]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size

Immunocytochemistry/ Immunofluorescence - Anti-MRP2 antibody [EPR10998] (AB172630)

  • 0
  • 0
Primary Antibodies

AB15580

Anti-Ki67 antibody

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (AB15580)

  • 5
  • 230
Primary Antibodies

AB109012

Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker

KO Validated|RabMAb|Recombinant|Lab Essentials

Western blot - Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker (AB109012)

  • 5
  • 15
Primary Antibodies

AB18207

Anti-beta III Tubulin antibody - Neuronal Marker

BOND RX™ Validated|Lab Essentials|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-beta III Tubulin antibody - Neuronal Marker (AB18207)

  • 5
  • 67
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

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Properties and storage information

Gene name
ABCC2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

MRP2 also known as ABCC2 or MRP-2 transporter is an ATP-binding cassette transporter. This protein with a molecular mass of about 190 kDa is widely expressed in tissues such as the liver kidney and intestine. MRP2 plays a role in drug and metabolite transport moving various organic anions across cellular membranes. It locates to the canalicular membrane of hepatocytes where it functions in secreting conjugated bilirubin and drugs into bile.
Biological function summary

MRP2 contributes to cellular detoxification processes. It does not form part of a larger protein complex but works as an independent entity. MRP2 protein actively transports conjugated substances out of cells protecting tissues from potential damage from toxins drugs and other organic anions. This transporter mediates the excretion of glucuronide sulfate and glutathione conjugates playing an important role in phase III of drug metabolism.

Pathways

MRP2 transporter is integral to xenobiotic metabolism. It interconnects with the conjugation pathways that prepare compounds for excretion. The pathway functions with cytochrome P450 enzymes and other related transport proteins like MDR1 facilitating the secretion of detoxified metabolites. MRP2 activity influences pharmacokinetics and pharmacodynamics by modulating drug disposition and excretion.

MRP2 is associated with Dubin-Johnson syndrome and cholestasis. Dubin-Johnson syndrome arises from mutations in the ABCC2 gene leading to defective bilirubin excretion and jaundice. Cholestasis involves impaired bile excretion where MRP2 and possibly related proteins like BSEP play roles in bile salt export. Understanding MRP2 functioning aids in assessing risk and treatment strategies for these conditions.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information ABCC2
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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