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AB261757

Human COIL (Coilin) knockout HeLa cell line

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COIL 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: 10 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human COIL (Coilin) knockout HeLa cell line (AB261757)
  • WB

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Western blot - Human COIL (Coilin) knockout HeLa cell line (AB261757)

Lanes 1-3 : Merged signal (red and green). Green - ab87913 observed at 75 kDa. Red - loading control ab181602 observed at 37 kDa.

ab87913 Anti-Coilin antibody [IH10] was shown to specifically react with Coilin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261757 (knockout cell lysate ab257251) was used. Wild-type and Coilin knockout samples were subjected to SDS-PAGE. ab87913 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Coilin antibody [IH10] (<a href='/en-us/products/primary-antibodies/coilin-antibody-ih10-ab87913'>ab87913</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

COIL knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human COIL (Coilin) knockout HeLa cell line (ab261757)

Lane 3:

Jurkat cell lysate at 20 µg

Predicted band size: 62 kDa

Observed band size: 75 kDa

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Western blot - Human COIL (Coilin) knockout HeLa cell line (AB261757)
  • WB

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Western blot - Human COIL (Coilin) knockout HeLa cell line (AB261757)

Western blot : Anti-COIL antibody [Pdelta] (ab11822) staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab11822 was shown to bind specifically to COIL. A band was observed at 68 kDa in wild-type HeLa cell lysates with no signal observed at this size in COIL knockout cell line. To generate this image, wild-type and COIL knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Coilin antibody [Pdelta] (<a href='/en-us/products/primary-antibodies/coilin-antibody-pdelta-ab11822'>ab11822</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lanes 1 - 4:

Western blot - Human COIL (Coilin) knockout HeLa cell line (ab261757)

Lane 2:

COIL knockout HeLa cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 68 kDa

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Western blot - Human COIL (Coilin) knockout HeLa cell line (AB261757)
  • WB

Lab

Western blot - Human COIL (Coilin) knockout HeLa cell line (AB261757)

Western blot : Anti-COIL antibody (ab210785) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab210785 was shown to bind specifically to COIL. A band was observed at 68 kDa in wild-type HeLa cell lysates with no signal observed at this size in COIL knockout cell line. To generate this image, wild-type and COIL knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Coilin antibody (<a href='/en-us/products/primary-antibodies/coilin-antibody-ab210785'>ab210785</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lanes 1 - 4:

Western blot - Human COIL (Coilin) knockout HeLa cell line (ab261757)

Lane 2:

COIL knockout HeLa cell lysate at 20 µg

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

SH-SY5Y cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 68 kDa

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Sanger Sequencing - Human COIL (Coilin) knockout HeLa cell line (AB261757)
  • Sanger seq

Unknown

Sanger Sequencing - Human COIL (Coilin) knockout HeLa cell line (AB261757)

Homozygous : 10 bp deletion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 10 bp deletion in exon 1

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB210785

Anti-Coilin antibody

KO Validated

Western blot - Anti-Coilin antibody (AB210785)

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  • 0
Cell Lines & Lysates

AB255444

Human VDAC1 (Porin) knockout HEK-293T cell line

Advanced Validation

Western blot - Human VDAC1 (Porin) knockout HEK-293T cell line (AB255444)

  • 0
  • 0
Primary Antibodies

AB179483

Anti-HIF-1 alpha antibody [EPR16897]

RabMAb|Advanced Validation|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] (AB179483)

  • 4
  • 9
Cell Lines & Lysates

AB255401

Human L1CAM knockout HeLa cell line

Advanced Validation

Immunocytochemistry/ Immunofluorescence - Human L1CAM knockout HeLa cell line (AB255401)

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  • 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
COIL
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.

Coilin also known as Coilin protein or its gene name COIL is a nuclear phosphoprotein with a mass of approximately 62 kDa. It plays a mechanical role in the formation and maintenance of Cajal bodies which are subnuclear structures involved in various RNA processes. Coilin is mainly expressed in the nucleus of eukaryotic cells including a high expression in HeLa cells where its localization to Cajal bodies can be easily studied with tools like Alexa Fluor 488. Occasionally references may be made to terms like 'HeLa coil' which pertain to specific structural features noted in these contexts.
Biological function summary

This protein facilitates the organization and biogenesis of nuclear bodies associated with RNA processing and small nuclear ribonucleoproteins (snRNPs) assembly. Coilin operates as part of a larger complex involving several other proteins that interact within Cajal bodies. These nuclear bodies serve as sites where components necessary for RNA processing are concentrated. Coilin aids in the trafficking and modification of snRNP elements which are essential for pre-mRNA splicing.

Pathways

Coilin affects RNA metabolism and processing notably within the spliceosome assembly pathway and in snRNP biogenesis. The pathways include interactions with proteins such as SMN (Survival of Motor Neuron) protein which links to spinal muscular atrophy-related processes. Additionally Coilin contributes to the broader framework of nucleic acid processing systems impacting the outcome of RNA transcription and modification through its regulatory roles.

Coilin shows connections to neurological conditions and cancer. For instance alterations or dysregulation in Coilin expression can correlate with spinal muscular atrophy where it associates with the SMN protein if there are disruptions in Cajal body integrity. Furthermore links exist with certain cancers where the mismanagement of RNA processes influenced by Coilin and its interaction partners like PDEdelta can lead to aberrant cell behaviors and tumor progression. Coilin's involvement in such diseases makes it an appealing target for therapeutic exploration reflected in ongoing cancer research and therapeutic developments.
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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 COIL
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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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