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AB267322

Human CAMK2D (CaMKII delta) knockout HEK-293T cell line

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CAMK2D KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 5 and 1 bp insertion in exon 5. 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 CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)
  • WB

Lab

Western blot - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)

Lanes 1- 2 : Merged signal (red and green). Green - ab181052 observed at 50 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab181052 was shown to react with CaM-kinase II in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab267322 (knockout cell lysate ab257376) was used. Wild-type HEK-293T and CAMK2D knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab181052 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-CaMKII delta antibody [EPR13095] (<a href='/en-us/products/primary-antibodies/camkii-delta-antibody-epr13095-ab181052'>ab181052</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

CAMK2D knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (ab267322)

Predicted band size: 56 kDa

Observed band size: 50 kDa

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Sanger Sequencing - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)

Allele-1 : 14 bp deletion in exon5

Cell Culture - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)
  • Cell Culture

Unknown

Cell Culture - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)

Representative images of CAMK2D knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.

Sanger Sequencing - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAMK2D (CaMKII delta) knockout HEK-293T cell line (AB267322)

Allele-2 : 1 bp insertion in exon 5.

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, 14 bp deletion in exon 5 and 1 bp insertion in exon 5

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Complementary Products

Primary Antibodies

AB181052

Anti-CaMKII delta antibody [EPR13095]

KO Validated|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaMKII delta antibody [EPR13095] (AB181052)

  • 5
  • 1
Primary Antibodies

AB320639

Anti-CAMK2B/2D/2G (phospho T287) + CAMK2A (phospho T286) antibody [EPR28075-37] - BSA and Azide free

Recombinant|RabMAb

Western blot - Anti-CAMK2B/2D/2G (phospho T287) + CAMK2A (phospho T286) antibody [EPR28075-37] - BSA and Azide free (AB320639)

  • 0
  • 0
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Primary Antibodies

AB124880

Anti-CaMKII alpha (phospho T286) antibody [EPR1830]

BOND RX™ Validated|RabMAb|Recombinant|Trial Size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CaMKII alpha (phospho T286) antibody [EPR1830] (AB124880)

  • 5
  • 1
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Reactivity data

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

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What's included?

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Properties and storage information

Gene name
CAMK2D
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
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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.

CaMKII delta also known as CaMKIIΔ or CaMK2D is a member of the CaMKII family of proteins recognized for its role as a serine/threonine protein kinase. This protein has a molecular mass of approximately 55 kDa. It is highly expressed in the brain and the heart where it plays essential roles in cellular functions. The domain structure of CaMKII delta allows for calcium/calmodulin activation which is important for its activity in phosphorylating substrates. This protein can bind to other intracellular molecules and is sensitive to regulation by its surrounding environment.
Biological function summary

CaMKII delta participates in several critical processes such as calcium signaling and regulation of gene expression. This protein influences synaptic plasticity muscle contraction and cell cycle control. It forms part of larger complexes interacting with other proteins and playing a role in adjusting cellular responses to various stimuli. It is also subject to inhibition with the compound Bisindolylmaleimide II known to influence its activity. Understanding the biological roles of CaMKII delta allows for insights into its effects on cellular mechanisms especially in neuronal and muscular tissues.

Pathways

CaMKII delta is significant in pathways like the calcium signaling pathway and the PI3K/Akt pathway. It serves as a modulator allowing the integration of various signals which is necessary for maintaining cellular homeostasis. CaMKII delta interacts with proteins such as protein kinase C (PKC) inhibitors affecting cellular signaling outputs. Its role in these pathways highlights the importance of CaMKII delta in regulating processes critical to cell survival growth and adaptation.

CaMKII delta is associated with conditions such as cardiac hypertrophy and Alzheimer's disease. These diseases often involve dysregulation of calcium signaling where CaMKII delta plays a part. CaMKII delta's association with proteins like PKC and their dysregulation contributes to the pathogenesis observed in these conditions. Understanding these interactions can aid in developing therapeutic strategies focused on modulating CaMKII delta activity to mitigate disease progression.
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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 CAMK2D
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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'.

For licensing inquiries, please contact partnerships@abcam.com