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AB265285

Human PUM1 (Pumilio 1) knockout HeLa cell line

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PUM1 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 121 bp insertion in exon 2 and 1 bp deletion in exon 2.

View Alternative Names

HsPUM, KIAA0099, PUM L1, PUM1_HUMAN, PUMH, PUMH 1, PUML 1, PUML1, Pumilio homolog 1, Pumilio-1, pumilio homolog 1 (Drosophila)

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Sanger Sequencing - Human PUM1 (Pumilio 1) knockout HeLa cell line (AB265285)
  • Sanger seq

Unknown

Sanger Sequencing - Human PUM1 (Pumilio 1) knockout HeLa cell line (AB265285)

Allele-2 : 121 bp insertion in exon 2.

Sanger Sequencing - Human PUM1 (Pumilio 1) knockout HeLa cell line (AB265285)
  • Sanger seq

Unknown

Sanger Sequencing - Human PUM1 (Pumilio 1) knockout HeLa cell line (AB265285)

Allele-1 : 1 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, 121 bp insertion in exon 2 and 1 bp deletion in exon 2

Antibiotic resistance

Puromycin 1µg/mL

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
PUM1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
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.

Pumilio 1 also known as PUM1 is a human protein coding for RNA-binding protein involved in the translational regulation of mRNA. It has a molecular weight of approximately 120 kDa. PUM1 expresses in various tissues with higher concentrations in brain placenta and testis. This protein contains a distinct Pumilio Homology Domain which allows it to specifically recognize and bind to a conserved sequence motif known as Pumilio Response Element (PRE) in target mRNAs.
Biological function summary

PUM1 is an important regulator of gene expression. It functions by binding to the 3' untranslated regions of target mRNAs leading to repression of translation and in some cases RNA decay. PUM1 operates within larger ribonucleoprotein complexes collaborating with other regulatory proteins to fine-tune the stability and translation of specific transcripts. This regulation is vital for processes like neuronal development stem cell maintenance and germ cell development.

Pathways

The influence of PUM1 extends to key biological networks such as the cell cycle and long-term potentiation pathways. It interacts with proteins like DDX6 which synergize to mediate post-transcriptional regulation as part of the mRNA decay pathway. Through its role in these pathways PUM1 helps ensure precise control of gene expression necessary for cellular homeostasis and adaptive responses to environmental changes.

The dysfunction of PUM1 is linked to neurological and reproductive conditions. Mutations or alterations in PUM1 expression have associations with spinocerebellar ataxia and gonadal dysgenesis. In these contexts PUM1 aberrations may also involve interactions with proteins such as FMRP known for its roles in neurological development and reproductive processes pointing to a complex network of genetic regulation and disease manifestation.

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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