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AB266368

Human UFSP2 knockout HEK-293T cell line

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UFSP2 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 6. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
3 Images
Western blot - Human UFSP2 knockout HEK-293T cell line (AB266368)
  • WB

Lab

Western blot - Human UFSP2 knockout HEK-293T cell line (AB266368)

Lanes 1-3 : Merged signal (red and green). Green - ab185965 observed at 53 kDa. Red - loading control ab8245 observed at 36 kDa.

ab185965 Anti-UFSP2 antibody [EPR13424] 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 (ab266368)

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 line (AB266368)
  • WB

Lab

Western blot - Human UFSP2 knockout HEK-293T cell line (AB266368)

Lanes 1-3 : Merged signal (red and green). Green - ab192597 observed at 53 kDa. Red - loading control ab8245 observed at 36 kDa.

ab192597 Anti-UFSP2 antibody [EP13424-49] 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 (ab266368)

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 line (AB266368)
  • Sanger seq

Unknown

Sanger Sequencing - Human UFSP2 knockout HEK-293T cell line (AB266368)

Homozygous : 1 bp deletion in exon6

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 6

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Complementary Products

Primary Antibodies

AB185965

Anti-UFSP2 antibody [EPR13424]

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-UFSP2 antibody [EPR13424] (AB185965)

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  • 0
Proteins & Peptides

AB313361

Recombinant Human WFDC2 Protein (His tag)

Mass Spectrometry - Recombinant Human WFDC2 Protein (His tag) (AB313361)

  • 0
  • 0
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
Proteins & Peptides

AB95933

Recombinant human XIAP protein

Functional Studies - Recombinant human XIAP protein (AB95933)

  • 0
  • 0
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Reactivity data

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Product details

We will provide viable cells that proliferate on revival.

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
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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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Target data

See full target information UFSP2
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
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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