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AB266086

Human CELF1 (CUG-BP1) knockout HEK-293T cell line

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CELF1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 2 and 5 bp deletion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Western blot - Human CELF1 (CUG-BP1) knockout HEK-293T cell line (AB266086)
  • WB

Lab

Western blot - Human CELF1 (CUG-BP1) knockout HEK-293T cell line (AB266086)

Lanes 1-4 : Merged signal (red and green). Green - ab129115 observed at 52 kDa. Red - loading control ab8245 observed at 36 kDa.

ab129115 Anti-CUG-BP1 antibody [EPR8298(B)] was shown to specifically react with CUG-BP1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266086 (knockout cell lysate ab257390) was used. Wild-type and CUG-BP1 knockout samples were subjected to SDS-PAGE. ab129115 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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-CUG-BP1 antibody [EPR8298(B)] (<a href='/en-us/products/primary-antibodies/cug-bp1-antibody-epr8298b-ab129115'>ab129115</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CELF1 (CUG-BP1) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-celf1-cug-bp1-knockout-hek-293t-cell-lysate-ab257390'>ab257390</a>) at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Human brain tissue 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: 52 kDa

Observed band size: 52 kDa

false

Sanger Sequencing - Human CELF1 (CUG-BP1) knockout HEK-293T cell line (AB266086)
  • Sanger seq

Unknown

Sanger Sequencing - Human CELF1 (CUG-BP1) knockout HEK-293T cell line (AB266086)

Allele-1 : 5 bp deletion in exon 2

Sanger Sequencing - Human CELF1 (CUG-BP1) knockout HEK-293T cell line (AB266086)
  • Sanger seq

Unknown

Sanger Sequencing - Human CELF1 (CUG-BP1) knockout HEK-293T cell line (AB266086)

Allele-2 : 4 bp deletion in exon 2.

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, 4 bp deletion in exon 2 and 5 bp deletion in exon 2

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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
CELF1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Quality control - STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Supplementary information

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

CUG-BP1 also known as CELF1 belongs to the CELF family of RNA-binding proteins. It is a 52-kDa protein expressed across various tissues with noticeable levels in heart and skeletal muscle. CUG-BP1 regulates alternative splicing by binding to specific CUG triplet repeats in RNA molecules. This binding influences the processing of pre-mRNAs impacting gene expression at the post-transcriptional level.
Biological function summary

The regulation by CUG-BP1 affects both mRNA stability and translation. The protein plays an important role in managing the expression of genes involved in muscle differentiation and development. CUG-BP1 often interacts with other cellular machinery forming functional complexes to facilitate its role in RNA metabolism. Its capacity to bind CUG motifs on target RNA highlights its significance in cellular processes.

Pathways

CUG-BP1 participates in pathways influencing muscle formation and maintenance. It plays part in the regulation of alternative splicing processes critical for efficient functioning of the p38 MAPK pathway. This pathway connects CUG-BP1 to other RNA-binding proteins and kinases. By doing so it affects different transcriptional and post-transcriptional processes emphasizing its critical function in cellular maintenance.

CUG-BP1 has been linked to myotonic dystrophy type 1 (DM1) and certain cancers. In DM1 the protein becomes sequestered by expanded CUG repeats in RNA disrupting its normal function. This sequestration leads to misregulated splicing events affecting various cellular functions. CUG-BP1's interaction with other proteins such as MBNL1 further elucidates its role in disease pathogenesis highlighting its importance in understanding RNA-mediated disorders.
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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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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Product protocols

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Target data

See full target information CELF1

Alternative Names

Deadenylation factor CUG-BP, RNA-binding protein BRUNOL-2, CUGBP1, CUG-BP1, CUG-BP- and ETR-3-like factor 1, BRUNOL2, Bruno-like protein 2, CUG triplet repeat RNA-binding protein 1, Embryo deadenylation element-binding protein homolog, 50 kDa nuclear polyadenylated RNA-binding protein, CELF1, CUGBP Elav-like family member 1, CELF-1, EDEN-BP homolog, NAB50, CUGBP
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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

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