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PCBD1 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: Insertion of the selection cassette in exon 2.

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Images

Western blot - Human PCBD1 knockout HEK-293T cell line (AB266261), expandable thumbnail
  • Cell Culture - Human PCBD1 knockout HEK-293T cell line (AB266261), expandable thumbnail
  • Sanger Sequencing - Human PCBD1 knockout HEK-293T cell line (AB266261), expandable thumbnail

Key facts

Cell type
HEK-293T
Species or organism
Human
Tissue
Kidney
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2

Alternative names

4-alpha-hydroxy-tetrahydropterin dehydratase, 6 pyruvoyl tetrahydropterin synthase/dimerization cofactor of hepatocyte nuclear factor 1 alpha, 6 pyruvoyl tetrahydropterin synthase/dimerization cofactor of hepatocyte nuclear factor 1 alpha (TCF1), DCoH, Dimerization cofactor of HNF1, Dimerization cofactor of hepatic nuclear factor 1 alpha, Dimerization cofactor of hepatocyte nuclear factor 1-alpha, Dimerizing cofactor for HNF1, PCBD, PCD, PHS_HUMAN, Phenylalanine hydroxylase-stimulating protein, Pterin 4 alpha carbinolamine dehydratase/dimerization cofactor of hepatocyte nuclear factor 1 alpha, Pterin 4 alpha carbinolamine dehydratase/dimerization cofactor of hepatocyte nuclear factor 1 alpha (TCF1), Pterin carbinolamine dehydratase, Pterin-4-alpha-carbinolamine dehydratase

Recommended products

PCBD1 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: Insertion of the selection cassette in exon 2.

Key facts

Cell type
HEK-293T
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 2
Concentration
Loading...

Properties

Gene name
PCBD1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

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

Cell culture

Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female

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.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type HEK293T cell line (ab255449). 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

PCBD1 also known as pterin-4-alpha-carbinolamine dehydratase or DCOH is a bifunctional protein with a molecular weight of approximately 12 kDa. It can be found expressed in a variety of tissues with significant expression noted in the liver and kidney. PCBD1 serves a dual role functioning both in enzymatic activity and as a co-regulator in transcription processes. Its enzymatic role involves catalyzing the dehydration of 4-alpha-hydroxy-tetrahydrobiopterin (BH4) an essential cofactor in the synthesis of neurotransmitters.

Biological function summary

PCBD1 interacts with tetrahydrobiopterin metabolism participating in neurotransmitter synthesis through its dehydratase function. It also acts as a dimerization cofactor for HNF1 (HNF1A) a transcription factor critical in gene regulation. The PCBD1 protein forms dimers or tetramers indicating its capability to participate in complex formation which enhances transcriptional regulation. This regulatory activity affects several genes involved in hepatic metabolism and development.

Pathways

PCBD1 primarily influences the tetrahydrobiopterin biosynthesis pathway and the regulation of gene expression pathway via HNF1A. By facilitating neurotransmitter production it indirectly plays a role in neurological pathways specifically those related to dopamine and serotonin synthesis. Through its transcriptional co-regulation with HNF1A PCBD1 impacts glucose metabolism and pancreatic functions linking it to metabolic pathways.

Associated diseases and disorders

Mutations or malfunctions of PCBD1 have associations with hyperphenylalaninemia a disorder affecting amino acid metabolism and with certain hyperinsulinemia forms influencing insulin regulation. Deficiencies in PCBD1 enzymatic function lead to altered levels of tetrahydrobiopterin while disruptions in its transcription co-regulation relate to impairments in glucose metabolism. Both HNF1A and PCBD1 mutations highlight their interconnected role in these conditions.

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