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AB257905

Human CSTB (Cystatin-B) knockout HEK-293T cell lysate

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CSTB KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.
2 Images
Western blot - Human CSTB (Cystatin-B) knockout HEK-293T cell lysate (AB257905)
  • WB

Unknown

Western blot - Human CSTB (Cystatin-B) knockout HEK-293T cell lysate (AB257905)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : CSTB knockout HEK293T cell lysate (20 ug)
Lane 3 : U87-MG cell lysate (20 ug)
Lane 4 : JAR cell lysate (20 ug)

ab92449 was shown to specifically react with Cystatin-B in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266321 (knockout cell lysate ab257905) was used. Wild-type and Cystatin-B knockout samples were subjected to SDS-PAGE. ab92449 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 CSTB (Cystatin-B) knockout HEK-293T cell lysate (AB257905)
  • Sanger seq

Unknown

Sanger Sequencing - Human CSTB (Cystatin-B) knockout HEK-293T cell lysate (AB257905)

Homozygous : Insertion of the selection cassette in exon 1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.

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

Cystatin-B also known as STEFIN-B is a protease inhibitor with a molecular mass of approximately 11 kilodaltons. It belongs to the family of cystatins which are expressed widely in human tissues with notable presence in the brain liver and kidneys. Cystatin-B acts by inhibiting cysteine proteases preventing them from degrading proteins indiscriminately. The inhibition is important for maintaining cellular protein balance and integrity.
Biological function summary

Cysteine protease inhibition by Cystatin-B protects cells from uncontrolled proteolysis which can lead to tissue damage. This protein plays an important role in the regulation of various cellular processes including apoptosis phagocytosis and the response to oxidative stress. It does not form part of a larger complex but interacts directly with its target proteases to exert its inhibitory effects.

Pathways

Cystatin-B influences proteolytic pathways by modulating the activity of cysteine proteases such as cathepsins. By inhibiting these enzymes Cystatin-B participates in the regulation of the lysosomal degradation pathway and apoptotic pathways which are important for cellular turnover and programmed cell death. Cathepsins particularly cathepsin B and L are related proteins that are significantly modulated through their interaction with Cystatin-B.

Mutations or dysregulation of Cystatin-B are associated with progressive myoclonus epilepsy and certain neurodegenerative diseases. The lack of functional Cystatin-B leads to excessive activity of proteases like cathepsin B resulting in increased proteolysis and neuronal damage. In some cases this can also connect to disorders involving oxidative stress exacerbating the detrimental cycle of cellular injury.

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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