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AB265959

Human PPP1R15B knockout HeLa cell line

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PPP1R15B KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

View Alternative Names

CREP, FLJ14744, Protein phosphatase 1, regulatory (inhibitor) subunit 15B, Protein phosphatase 1, regulatory subunit 15B

1 Images
Sanger Sequencing - Human PPP1R15B knockout HeLa cell line (AB265959)
  • Sanger seq

Unknown

Sanger Sequencing - Human PPP1R15B knockout HeLa cell line (AB265959)

Homozygous : 1 bp insertion in exon 1.

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: 1 bp insertion in exon 1

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, 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
PPP1R15B
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.

PPP1R15B also known as GADD34 is a regulatory subunit of protein phosphatase 1 (PP1). It functions mechanically by associating with PP1 to regulate the dephosphorylation of the eukaryotic initiation factor 2 alpha (eIF2α). This interaction is essential for controlling protein synthesis especially during stress conditions. The molecular mass of PPP1R15B is approximately 74 kDa. Expression of PPP1R15B occurs in various tissues with high levels in brain liver and kidney tissues.
Biological function summary

The regulatory action of PPP1R15B on eIF2α impacts protein synthesis by reversing the phosphorylation which occurs in response to cellular stress. It functions as part of a multi-protein complex that includes PP1 and possibly other proteins involved in cellular stress responses. As a result it helps in the recovery of protein translation during periods of endoplasmic reticulum stress contributing to cell survival pathways under transient stress conditions.

Pathways

PPP1R15B plays a significant role in the integrated stress response pathway by modulating protein synthesis. It participates in the unfolded protein response (UPR) which is important when cells encounter endoplasmic reticulum stress. Through its interaction with eIF2α and other associated components like PP1 PPP1R15B facilitates cellular adaptation to changing environmental stressors.

PPP1R15B has been implicated in diseases such as neurodegenerative disorders and certain types of cancer. Deregulation of its function can lead to aberrant protein synthesis control which results in pathological outcomes. In neurodegenerative diseases its interaction with proteins like eIF2α is significant affecting neuronal cell survival under stress. In cancer altered expression or function of PPP1R15B may contribute to tumor progression due to its role in managing cellular stress responses and apoptosis.

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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