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AB264712

Human CAMK2G (CamKII gamma) knockout HeLa cell line

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CAMK2G KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and 2 bp deletion in exon 4.

View Alternative Names

CAMK, CAMK II, CAMK2G, CAMKG, CaM kinase II, CaM kinase II gamma chain, CaM kinase II subunit gamma, CaMK II subunit gamma

4 Images
Western blot - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)
  • WB

Lab

Western blot - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)

Lanes 1-3 : Merged signal (red and green). Green - ab201966 observed at 63 kDa. Red - loading control ab181602 observed at 37 kDa.

ab201966 Anti-CamKII gamma antibody [8G10C1] was shown to specifically react with CamKII gamma in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264712 (knockout cell lysate ab258340) was used. Wild-type and CamKII gamma knockout samples were subjected to SDS-PAGE. ab201966 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-CamKII gamma antibody [8G10C1] (<a href='/en-us/products/primary-antibodies/camkii-gamma-antibody-8g10c1-ab201966'>ab201966</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CAMK2G knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CAMK2G (CamKII gamma) knockout HeLa cell line (ab264712)

Lane 3:

T-47D cell lysate at 20 µg

Predicted band size: 63 kDa

Observed band size: 63 kDa

false

Sanger Sequencing - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)

Allele-2 : 1 bp deletion in exon 4.

Sanger Sequencing - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)

Allele-1 : 2 bp deletion in exon 4.

Sanger Sequencing - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAMK2G (CamKII gamma) knockout HeLa cell line (AB264712)

Allele-3 : 1 bp insertion in exon 4.

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, 1 bp deletion in exon 4 and 1 bp insertion in exon 4 and 2 bp deletion in exon 4

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

Recommended control: Human wild-type HeLa cell line (ab255928). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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
CAMK2G
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
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

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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