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AB257569

Human OTUB1 knockout HEK-293T cell lysate

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OTUB1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

View Alternative Names

Deubiquitinating enzyme OTUB1, HSPC263, OTB1, OTU domain, ubiquitin aldehyde binding 1, OTU domain-containing Ubal-binding protein 1, OTU domain-containing ubiquitin aldehyde-binding protein 1, OTU-domain Ubal-binding 1, OTUB1_HUMAN, Otubain-1, Ubiquitin thioesterase OTUB1, Ubiquitin-specific-processing protease OTUB1, hOTU1, ubiquitin-specific protease otubain 1

4 Images
Western blot - Human OTUB1 knockout HEK-293T cell lysate (AB257569)
  • WB

Lab

Western blot - Human OTUB1 knockout HEK-293T cell lysate (AB257569)

Lane 1 : Wild-type HEK-293T cell lysate (20 μg)

Lane 2 : OTUB1 knockout HEK-293T cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab101471 observed at 130 kDa. Red - loading control ab8245 observed at 37 kDa.

ab101471 Anti-OTUB1 antibody was shown to specifically react with OTUB1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266551 (knockout cell lysate ab257569) was used. Wild-type and OTUB1 knockout samples were subjected to SDS-PAGE. ab101471 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 ug/ml 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.

All lanes:

Western blot - Anti-OTUB1 antibody (<a href='/en-us/products/primary-antibodies/otub1-antibody-ab101471'>ab101471</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

OTUB1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human OTUB1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-otub1-knockout-hek-293t-cell-line-ab266551'>ab266551</a>)

Predicted band size: 31 kDa

Observed band size: 130 kDa

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Western blot - Human OTUB1 knockout HEK-293T cell lysate (AB257569)
  • WB

Lab

Western blot - Human OTUB1 knockout HEK-293T cell lysate (AB257569)

Lane 1 : Wild-type HEK-293T cell lysate (20 μg)

Lane 2 : OTUB1 knockout HEK-293T cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab175200 observed at 31 kDa. Red - loading control ab8245 observed at 37 kDa.

ab175200 Anti-OTUB1 antibody [EPR13028(B)] was shown to specifically react with OTUB1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266551 (knockout cell lysate ab257569) was used. Wild-type and OTUB1 knockout samples were subjected to SDS-PAGE. ab175200 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.

All lanes:

Western blot - Anti-OTUB1 antibody [EPR13028(B)] (<a href='/en-us/products/primary-antibodies/otub1-antibody-epr13028b-ab175200'>ab175200</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

OTUB1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human OTUB1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-otub1-knockout-hek-293t-cell-line-ab266551'>ab266551</a>)

Predicted band size: 31 kDa

Observed band size: 130 kDa,31 kDa

false

Sanger Sequencing - Human OTUB1 knockout HEK-293T cell lysate (AB257569)
  • Sanger seq

Unknown

Sanger Sequencing - Human OTUB1 knockout HEK-293T cell lysate (AB257569)

Allele-2 : Insertion of the selection cassette in exon 1

Sanger Sequencing - Human OTUB1 knockout HEK-293T cell lysate (AB257569)
  • Sanger seq

Unknown

Sanger Sequencing - Human OTUB1 knockout HEK-293T cell lysate (AB257569)

Allele-1 : 19 bp deletion in exon 1

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, 19 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.

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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.

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What's included?

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Properties and storage information

Gene name
OTUB1
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
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Supplementary information

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

The OTUB1 protein also known as OTU domain-containing ubiquitin aldehyde-binding protein 1 plays an important role in the removal of ubiquitin from proteins. This process is important for regulating protein stability and function within the cell. OTUB1 has a mass of approximately 32 kDa and is expressed ubiquitously across various tissues indicating its essential nature in cellular functions.
Biological function summary

OTUB1 functions as a deubiquitinating enzyme that cleaves ubiquitin from substrates influencing various cellular processes. It acts independently or as part of larger protein complexes. By controlling protein degradation pathways OTUB1 impacts protein turnover and cellular homeostasis. Interactions with other proteins allow OTUB1 to exert influence on different signaling pathways and cellular stress responses.

Pathways

OTUB1 holds a significant position in the regulation of the ubiquitin-proteasome system and DNA damage repair pathway. It closely interacts with proteins such as UBE2N affecting the DNA damage response. Additionally OTUB1 regulates processes through modulating pathways like NF-kB by interacting with and stabilizing specific protein factors involved.

OTUB1’s function connects it to various pathological states like cancer and neurodegenerative diseases. Aberrant expression or mutations in OTUB1 can lead to disruptions in cell cycle regulation and apoptosis contributing to oncogenesis. In the context of neurodegenerative diseases OTUB1 is related to tau stabilization with proteins such as HSP90 playing a role in the same regulatory mechanisms highlighting its relevance in these disorders.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Western blot - Anti-OTUB1 antibody [EPR13028(B)] (AB175200)

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Product promise

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