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AB264735

Human KCNJ11 (Kir6.2/BIR) knockout HeLa cell line

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KCNJ11 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 25 bp deletion in exon 2.

View Alternative Names

ATP sensitive inward rectifier potassium channel 11, BIR, Beta cell inward rectifier subunit, HHF 2, IKATP, IRK 11, Inward rectifier K(+) channel Kir6.2, Inwardly rectifying potassium channel KIR6.2, KCNJ11, Kir 6.2, MGC133230, PHHI, Potassium channel inwardly rectifing subfamily J member 11, Potassium channel, inwardly rectifying subfamily J member 11, Potassium inwardly rectifying channel J11, TNDM 3

1 Images
Sanger Sequencing - Human KCNJ11 (Kir6.2/BIR) knockout HeLa cell line (AB264735)
  • Sanger seq

Unknown

Sanger Sequencing - Human KCNJ11 (Kir6.2/BIR) knockout HeLa cell line (AB264735)

Homozygous : 25 bp deletion in exon 2.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 25 bp deletion in exon 2

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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.

What's included?

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Properties and storage information

Gene name
KCNJ11
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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.

Kir6.2 also known by alternate names KCNJ11 and 2BIR is part of the ATP-sensitive potassium channel (K-ATP channel) complex specifically the pore-forming subunit. This protein has a molecular mass of approximately 43 kDa. Expression of Kir6.2 occurs in various tissues most notably in pancreatic beta cells cardiac muscle and neuronal tissues. These channels play an important role in cell membrane potential regulation and excitability modulating insulin release and muscle contraction by responding to intracellular levels of ATP and ADP.
Biological function summary

Kir6.2 functions by coupling cellular metabolic states to electrical activity via its role in the K-ATP channel complex. This complex integrates Kir6.2 with the sulfonylurea receptor SUR1 or SUR2 forming an important connection between cellular metabolism and membrane excitability. Kir6.2 helps manage glucose-induced insulin secretion in pancreatic beta cells contributing to the temporal burst of insulin after meals. Its function in neurons and muscle fibers includes balancing cellular energy levels and physiological processes such as neurotransmitter release and muscle contraction.

Pathways

Kir6.2 plays a significant role in insulin secretion and cardiac muscle contraction pathways. It interacts closely with various proteins including SUR1 in the insulin release pathway and SUR2 in cardiovascular regulation. The activity of Kir6.2 links it with processes such as glucose-stimulated insulin secretion where its modulation significantly impacts the entry of calcium ions through voltage-dependent calcium channels further altering the exocytosis of insulin granules. The reactivity to intracellular ATP levels means Kir6.2 acts as a metabolic sensor influencing these pathways accordingly.

Kir6.2 is linked to conditions such as neonatal diabetes mellitus and congenital hyperinsulinism. Mutations in the gene encoding Kir6.2 can disrupt normal K-ATP channel functioning leading to these diseases. In neonatal diabetes Kir6.2 alterations impair insulin secretion due to disrupted ATP binding or channel closure while in congenital hyperinsulinism dysfunction of Kir6.2 in hyperactive channels results in excessive insulin production. This exposes strong connections with proteins like insulin within these conditions highlighting its important influence on glucose metabolism and energy homeostasis.

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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