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AB265097

Human CRK (Crk p38) knockout HeLa cell line

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CRK 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 insertion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

View Alternative Names

Adapter molecule crk, Avian sarcoma virus CT10 (v crk) oncogene homolog, CRK isoform 2, CRK isoform II, CRKII, CRK_HUMAN, FLJ38130, OTTHUMP00000115366, OTTHUMP00000198330, P38, Proto-oncogene C-crk, v crk avian sarcoma virus CT10 oncogene homolog, v crk sarcoma virus CT10 oncogene homolog, v crk sarcoma virus CT10 oncogene homolog (avian)

3 Images
Sanger Sequencing - Human CRK (Crk p38) knockout HeLa cell line (AB265097)
  • Sanger seq

Unknown

Sanger Sequencing - Human CRK (Crk p38) knockout HeLa cell line (AB265097)

Homozygous : 1 bp insertion in exon 1.

Western blot - Human CRK (Crk p38) knockout HeLa cell line (AB265097)
  • WB

Lab

Western blot - Human CRK (Crk p38) knockout HeLa cell line (AB265097)

Western blot : Anti-Crk p38 antibody [22/Crk] ab300630 staining at 1/1000 dilution, shown in green; Rabbit anti alpha Tubulin ab52866 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 35, 24 kDa in Wild-type HeLa cell lysates with no signal observed at this size in CRK knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 800CW and Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Crk p38 antibody [22/Crk] (<a href='/en-us/products/primary-antibodies/crk-p38-antibody-22-crk-ab300630'>ab300630</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa whole cell lysate at 20 µg

Lane 2:

Western blot - Human CRK (Crk p38) knockout HeLa cell line (ab265097) at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Lane 4:

K562 whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 34 kDa

Observed band size: 35 kDa,24 kDa

false

Western blot - Human CRK (Crk p38) knockout HeLa cell line (AB265097)
  • WB

Lab

Western blot - Human CRK (Crk p38) knockout HeLa cell line (AB265097)

Western blot : Anti-Crk p38 antibody [EP242Y] ab45136 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin ab7291 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 35 kDa in Wild-type HeLa cell lysates with no signal observed at this size in CRK knockout HeLa cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Crk p38 antibody [EP242Y] (<a href='/en-us/products/primary-antibodies/crk-p38-antibody-ep242y-ab45136'>ab45136</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa whole cell lysate at 20 µg

Lane 2:

Western blot - Human CRK (Crk p38) knockout HeLa cell line (ab265097) at 20 µg

Lane 3:

HepG2 whole cell lysate at 20 µg

Lane 4:

K562 whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

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: 1 bp insertion in exon 1

Disease

Adenocarcinoma

Reactivity data

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

Product details

We will provide viable cells that proliferate on revival.

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.

What's included?

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Properties and storage information

Gene name
CRK
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

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.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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