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AB258095

Human PCBD1 knockout HEK-293T cell lysate

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PCBD1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon2.

View 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

2 Images
Western blot - Human PCBD1 knockout HEK-293T cell lysate (AB258095)
  • WB

Unknown

Western blot - Human PCBD1 knockout HEK-293T cell lysate (AB258095)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : PCBD1 knockout HEK293T cell lysate (20 ug)
Lane 3 : Caco-2 cell lysate (20 ug)
Lane 4 : Human ovary cancer tissue lysate (20 ug)

ab138518 was shown to specifically react with PCBD1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266261 (knockout cell lysate ab258095) was used. Wild-type and PCBD1 knockout samples were subjected to SDS-PAGE. ab138518 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 500 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.

false

Sanger Sequencing - Human PCBD1 knockout HEK-293T cell lysate (AB258095)
  • Sanger seq

Unknown

Sanger Sequencing - Human PCBD1 knockout HEK-293T cell lysate (AB258095)

Homozygous : Insertion of the selection cassette in exon2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon2.

Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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
PCBD1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C

Supplementary information

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

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.

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.

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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