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AB258082

Human NSDHL knockout HEK-293T cell lysate

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NSDHL KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 4.

View Alternative Names

H105E3, H105e3 protein, NAD(P) dependent steroid dehydrogenase like, NSDHL_HUMAN, Protein H105e3, SDR31E1, Short chain dehydrogenase/reductase family 31E member 1, Sterol 4 alpha carboxylate 3 dehydrogenase decarboxylating, Sterol-4-alpha-carboxylate 3-dehydrogenase, XAP104, decarboxylating

3 Images
Western blot - Human NSDHL knockout HEK-293T cell lysate (AB258082)
  • WB

Unknown

Western blot - Human NSDHL knockout HEK-293T cell lysate (AB258082)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : NSDHL knockout HEK293T cell lysate (20 ug)
Lane 3 : A431 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab199730 was shown to specifically react with NSDHL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266682 (knockout cell lysate ab258082) was used. Wild-type and NSDHL knockout samples were subjected to SDS-PAGE. ab199730 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4oC at 1 in 1000 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

Western blot - Human NSDHL knockout HEK-293T cell lysate (AB258082)
  • WB

Unknown

Western blot - Human NSDHL knockout HEK-293T cell lysate (AB258082)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : NSDHL knockout HEK293T cell lysate (20 ug)
Lane 3 : A431 cell lysate (20 ug)
Lane 4 : HeLa cell lysate (20 ug)

ab190353 was shown to specifically react with NSDHL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266682 (knockout cell lysate ab258082) was used. Wild-type and NSDHL knockout samples were subjected to SDS-PAGE. ab190353 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4oC at 1 in 1000 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 NSDHL knockout HEK-293T cell lysate (AB258082)
  • Sanger seq

Unknown

Sanger Sequencing - Human NSDHL knockout HEK-293T cell lysate (AB258082)

Homozygous : 1 bp deletion in exon 4

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, Homozygous: 1 bp deletion in exon 4.

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 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
NSDHL
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous
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.

NSDHL also known as NAD(P)H steroid dehydrogenase-like is an enzyme linked to the cholesterol biosynthesis pathway. It has a molecular weight of approximately 41 kDa. This enzyme localizes to the endoplasmic reticulum membrane and the peroxisomes. NSDHL shows expression in various tissues with higher levels in the liver adrenal glands and the brain. This distribution highlights its essential role in metabolic processes across different tissue types.
Biological function summary

NSDHL facilitates the removal of one hydrogen from NAD(P)H and adds it to the steroid precursor lathosterol an important intermediate in cholesterol biosynthesis. Although not part of a larger complex NSDHL performs significant enzymatic steps necessary for converting sterol intermediates. Its activity directly influences sterol composition vital for cell membrane integrity signaling and hormone synthesis.

Pathways

The enzyme plays a critical role in the cholesterol biosynthesis pathway specifically affecting the conversion of lanosterol into cholesterol. NSDHL operates in tandem with other enzymes like DHCR7 influencing the downstream process of sterol maturation. The precise coordination of these enzymes ensures effective cholesterol production essential for maintaining cellular cholesterol levels and systemic lipid homeostasis.

Malfunctions in NSDHL can lead to disorders like CHILD syndrome (Congenital Hemidysplasia with Ichthyosiform erythroderma and Limb Defects) and CK syndrome (Conradi-Hünermann-Happle syndrome). These conditions link to cholesterol biosynthesis disruption impacting skin and skeletal development. NSDHL interacts with the EBP protein in these pathways suggesting a relationship between enzyme activity and phenotypic expression in these disorders. Understanding NSDHL's function and regulation is key to recognizing its involvement in metabolic syndromes and potential therapeutic targets.

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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For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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