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AB266528

Human GNB2 knockout HEK-293T cell line

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GNB2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

G protein beta 2 subunit, G protein subunit beta-2, GBB2_HUMAN, Gnb2l1, Guanine nucleotide binding protein G I G S G T beta 2 subunit 2, Guanine nucleotide binding protein G protein beta polypeptide 2, Guanine nucleotide binding protein beta 2 subunit, Guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2, OTTHUMP00000174601, OTTHUMP00000174602, Receptor for activated C kinase, Receptor of activated protein kinase C 1, Signal transducing guanine nucleotide binding regulatory protein beta, Transducin beta chain 2

2 Images
Western blot - Human GNB2 knockout HEK-293T cell line (AB266528)
  • WB

Lab

Western blot - Human GNB2 knockout HEK-293T cell line (AB266528)

Lanes 1-4 : Merged signal (red and green). Green - ab108504 observed at 35 kDa. Red - loading control ab7291 observed at 50 kDa.

ab108504 Anti-GNB2 antibody [EP3262Y] was shown to specifically react with GNB2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266528 (knockout cell lysate ab257453) was used. Wild-type and GNB2 knockout samples were subjected to SDS-PAGE. ab108504 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2. 5 hours 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-GNB2 antibody [EP3262Y] (<a href='/en-us/products/primary-antibodies/gnb2-antibody-ep3262y-ab108504'>ab108504</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

GNB2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human GNB2 knockout HEK-293T cell line (ab266528)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

All lanes:

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 1/10000 dilution

Predicted band size: 37 kDa

Observed band size: 35 kDa

false

Sanger Sequencing - Human GNB2 knockout HEK-293T cell line (AB266528)
  • Sanger seq

Unknown

Sanger Sequencing - Human GNB2 knockout HEK-293T cell line (AB266528)

Homozygous : 1 bp insertion in exon2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2

Reactivity data

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

Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
GNB2
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

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.

GNB2 also known as Guanine Nucleotide Binding Protein Subunit Beta 2 is part of the G protein family. It has a molecular mass of approximately 38 kDa. GNB2 functions mechanically by acting as a modulator or transducer in various signaling pathways. This protein is expressed in a wide range of tissues including the brain heart and muscles. Its role involves interacting with alpha and gamma subunits to form a heterotrimeric G protein complex that is important for signal transduction.
Biological function summary

In the context of cellular processes GNB2 is involved in transmitting signals from activated G protein-coupled receptors to different intracellular effectors. It forms an essential part of the G protein complex which plays a role in facilitating the exchange of GDP for GTP on the alpha subunit. The activation of the G protein complex enables downstream signaling that governs multiple physiological responses such as sensory perception and cellular communication.

Pathways

GNB2 is heavily involved in pathways related to signal transduction and sensory perception. It plays a significant role within the GPCR (G protein-coupled receptor) signaling pathway an important pathway involved in transmitting signals from diverse stimuli outside a cell to its interior. In this context GNB2 interacts with related proteins such as GNAI2 contributing to the regulation of ion channels adenylyl cyclases and other effectors. This connectivity enables precise cellular responses to external signals.

Research indicates that GNB2 is associated with cardiac arrhythmias and neuropsychiatric disorders. It has connections to other proteins like GNAO1 in the context of these conditions. Mutations or dysregulation of GNB2 can disrupt normal signaling processes leading to these diseases. Understanding the role of GNB2 in these disorders highlights potential therapeutic targets for developing new interventions.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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