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AB262320

Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line

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ARRB2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 4.

View Alternative Names

ARB 2, ARR 2, ARRB2_HUMAN, Arrestin 3, Arrestin beta-2, BARR2, Beta-arrestin-2, DKFZp686L0365, HGNC:712

3 Images
Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (AB262320)
  • WB

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Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (AB262320)

Lanes 1 - 4 : Merged signal (red and green). Green - ab54790 observed at 50 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.

ab54790 was shown to react with Beta Arrestin 2 in wild-type HepG2 cells in Western blot with loss of signal observed in ARRB2 knockout cell line ab262320 (knockout cell lysate ab257283). Wild-type HepG2 and ARRB2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab54790 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (<a href='/en-us/products/primary-antibodies/beta-arrestin-2-antibody-3g1-ab54790'>ab54790</a>) at 1 µg/mL

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

ARRB2 knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (ab262320)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Human Lung tissue lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 50 kDa

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Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (AB262320)
  • WB

Lab

Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (AB262320)

Lanes 1- 4 : Merged signal (red and green). Green - ab54790 observed at 55 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

ab54790 was shown to react with ARRB2 in wild-type HepG2 cells in western blot. Loss of signal was observed when knockout cell line ab262320 (knockout cell lysate ab257283) was used. Wild-type HepG2 and ARRB2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab54790 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Beta Arrestin 2 antibody [3G1] (<a href='/en-us/products/primary-antibodies/beta-arrestin-2-antibody-3g1-ab54790'>ab54790</a>) at 1/500 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

ARRB2 knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (ab262320)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

K562 cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 55 kDa

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Sanger Sequencing - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (AB262320)
  • Sanger seq

Unknown

Sanger Sequencing - Human ARRB2 (Beta Arrestin 2) knockout Hep G2 cell line (AB262320)

Homozygous : 1 bp deletion in exon4

Key facts

Cell type

Hep G2

Species or organism

Human

Tissue

Liver

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 4

Antibiotic resistance

Puromycin 1µg/mL

Disease

Hepatocellular Carcinoma

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Reactivity data

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Product details

Recommended control: Human wild-type HepG2 cell line (ab257304). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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
ARRB2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.
Culture medium

MEM + 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.

Beta-arrestin 2 also known as arrb2 or b-arrestin mechanically functions to regulate G protein-coupled receptor (GPCR) signaling. This protein has a molecular mass of approximately 47 kDa and is ubiquitously expressed across various tissues including the brain lungs and heart. Beta-arrestin 2 works by binding to phosphorylated GPCRs facilitating receptor desensitization and endocytosis which stops the receptor's signaling activity and mediates receptor internalization.
Biological function summary

Beta-arrestin 2 plays an important role in mediating signal transduction pathways beyond GPCR desensitization including those involved in cell communication and cellular response regulation. Beta-arrestin 2 interacts with multiple signaling proteins to form complexes that facilitate signaling pathways notably influencing MAPK (Mitogen-Activated Protein Kinase) pathways. It provides a scaffold for downstream signaling molecules affecting cell signaling outcomes.

Pathways

Beta-arrestin 2 significantly participates in two important biological pathways: the MAPK pathway and the Wnt signaling pathway. Through these pathways beta-arrestin 2 interacts with signaling proteins such as ERK1/2 (Extracellular signal-Regulated Kinases) and dishevelled proteins. These interactions highlight beta-arrestin 2's ability to modulate signal strength and duration contributing to diverse cellular responses.

Beta-arrestin 2 is implicated in conditions like heart failure and certain cancers. The aberrant regulation of beta-arrestin 2-related pathways can lead to uncontrolled cell proliferation or inadequate heart responses. Additionally beta-arrestin 2 is connected to proteins like GPCR kinases in the context of heart failure and MAPK proteins within oncogenic processes underlining its potential as a therapeutic target.
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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 ARRB2
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