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AB257782

Human UFSP2 knockout HEK-293T cell lysate

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UFSP2 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon6.

View Alternative Names

C4orf20, Chromosome 4 open reading frame 20, FLJ11200, UFM1-specific peptidase 2, UFSP2_HUMAN, Ufm1-specific protease 2

3 Images
Western blot - Human UFSP2 knockout HEK-293T cell lysate (AB257782)
  • WB

Lab

Western blot - Human UFSP2 knockout HEK-293T cell lysate (AB257782)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : UFSP2 knockout HEK293T cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)

ab185965 was shown to specifically react with UFSP2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266368 (knockout cell lysate ab257782) was used. Wild-type and UFSP2 knockout samples were subjected to SDS-PAGE. ab185965 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-UFSP2 antibody [EPR13424] (<a href='/en-us/products/primary-antibodies/ufsp2-antibody-epr13424-ab185965'>ab185965</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

UFSP2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human UFSP2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ufsp2-knockout-hek-293t-cell-line-ab266368'>ab266368</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

Western blot - Human UFSP2 knockout HEK-293T cell lysate (AB257782)
  • WB

Lab

Western blot - Human UFSP2 knockout HEK-293T cell lysate (AB257782)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : UFSP2 knockout HEK293T cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)

ab192597 was shown to specifically react with UFSP2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266368 (knockout cell lysate ab257782) was used. Wild-type and UFSP2 knockout samples were subjected to SDS-PAGE. ab192597 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-UFSP2 antibody [EP13424-49] (<a href='/en-us/products/primary-antibodies/ufsp2-antibody-ep13424-49-ab192597'>ab192597</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

UFSP2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human UFSP2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ufsp2-knockout-hek-293t-cell-line-ab266368'>ab266368</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

Sanger Sequencing - Human UFSP2 knockout HEK-293T cell lysate (AB257782)
  • Sanger seq

Unknown

Sanger Sequencing - Human UFSP2 knockout HEK-293T cell lysate (AB257782)

Homozygous : 1 bp deletion in exon6

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, 1 bp deletion in exon6.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

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Properties and storage information

Gene name
UFSP2
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.

UFSP2 also known as Ubiquitin-Fold Modifier 1-Specific Peptidase 2 is a protein with a molecular mass of approximately 53 kDa. This protein functions as a cysteine protease involved in the processing and maturation of ubiquitin-fold modifier 1 (UFM1). UFSP2 cleaves precursor UFM1 to expose the glycine residue necessary for its conjugation to target proteins. Expression of UFSP2 occurs in various tissues but is particularly high in the testis brain and skeletal muscle indicating its roles in diverse biological functions.
Biological function summary

UFSP2 plays a significant role in the UFMylation pathway which is a post-translational modification process involving UFM1. This enzyme does not operate alone; it acts as part of a functional complex where its peptidase activity is essential for recycling UFM1 molecules after they detach from substrate proteins. The activation and deactivation of UFM1-modified proteins regulate several cellular processes such as protein translation cellular stress responses and maintenance of endoplasmic reticulum homeostasis.

Pathways

The UFSP2 protein contributes to the wider ubiquitin-like modification systems particularly impacting the protein quality control system and the endoplasmic reticulum-associated degradation (ERAD) pathway. It maintains protein folding and ensures degradation of misfolded proteins. UFSP2 interacts with other proteins such as UFL1 which acts as a specific E3 ligase for UFM1 establishing a cycle for efficient modification and de-modification in these pathways.

Alterations in UFSP2 function link to pathologies including autosomal-dominant osteosclerosis with the novel symptom of cranial sclerosis (ADOCS). Mutations in the UFSP2 gene can lead to defective bone formation due to dysregulation of protein homeostasis pathways. Furthermore anomalies in UFSP2 have associations with neurodegenerative diseases where it interacts with proteins involved in cellular stress responses. This highlights its importance in both skeletal and nervous system-related 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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