JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB265080

Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line

Be the first to review this product! Submit a review

|

(0 Publication)

CACNA2D1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon 11 and 14 bp deletion in exon 11 and 1 bp insertion in exon 11. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

CA2D1_HUMAN, CACN A2, CACNL2A, CCHL2A, Cacna2d1, Calcium channel L type alpha 2 polypeptide, Calcium channel voltage dependent alpha 2/delta subunit 1, Dihydropyridine receptor alpha 2 subunit, Dihydropyridine sensitive L type calcium channel alpha 2/delta subunit, Dihydropyridine sensitive L type calcium channel subunits alpha 2/delta, L type calcium channel subunit alpha 2, MHS 3, Voltage dependent calcium channel subunit alpha 2/delta 1, Voltage-dependent calcium channel subunit delta-1, Voltage-gated calcium channel subunit alpha-2/delta-1

5 Images
Western blot - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)
  • WB

Unknown

Western blot - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)

Lanes 1-2 : Merged signal (red and green). Green - ab2864 observed at 150 kDa. Red - loading control ab181602 observed at 37 kDa.

ab2864 Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [20A] was shown to specifically react with Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265080 (knockout cell lysate ab257374) was used. Wild-type and Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 knockout samples were subjected to SDS-PAGE. ab2864 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°CC 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [20A] (<a href='/en-us/products/unavailable/calcium-channel-l-type-dhpr-alpha-2-subunitcacna2d1-antibody-20a-ab2864'>ab2864</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CACNA2D1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (ab265080)

false

Sanger Sequencing - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)
  • Sanger seq

Unknown

Sanger Sequencing - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)

Allele-3 : 1 bp insertion in exon 11.

Sanger Sequencing - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)
  • Sanger seq

Unknown

Sanger Sequencing - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)

Allele-2 : 11 bp deletion in exon 11.

Sanger Sequencing - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)
  • Sanger seq

Lab

Sanger Sequencing - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)

Allele-1 : 14 bp deletion in exon 11.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line (AB265080)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type Hela ( tissue labeling Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 with ab313763 at 1/500 (1.046 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) Wild-type Hela cell pellet; no staining on (B) CACNA2D1 knockout HeLa cell pellet and (C) THP-1 cell pellet. The section was incubated with ab313763 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

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, 11 bp deletion in exon 11 and 14 bp deletion in exon 11 and 1 bp insertion in exon 11

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

style
left-column

Reactivity data

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

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.

style
left-column

What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab265080-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab255928 Human wild-type HeLa cell line", "number":"AB265080-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab265080 Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line", "number":"AB265080-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab265080-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab265080 Human CACNA2D1 (Calcium channel L type DHPR alpha 2 subunit) knockout HeLa cell line", "number":"AB265080-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
CACNA2D1
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
style
left-column

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.

style
left-column

Supplementary information

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

The Calcium channel L type DHPR alpha 2 subunit also known as CACNA2D1 or alpha-2-delta plays a significant role in cellular calcium regulation. This subunit typically part of the voltage-gated calcium channel complex allows calcium ions to enter the cells following electrical stimuli. With a mass of about 150 kDa CACNA2D1 expresses predominantly in skeletal and cardiac muscle tissues and the central nervous system. Its presence across different tissues suggests various physiological roles linking its mechanical function to electrical signaling and muscle contraction.
Biological function summary

CACNA2D1 aids in modulating neurotransmitter release and excitation-contraction coupling. This protein often integrates into the larger voltage-gated calcium channel complex working in concert with other subunits to optimize calcium flow. This particular function supports critical processes in muscle cells and neurons integrating signals in response to electrical impulses. The alpha-2-delta ligand poses significance here as well often used to modify the channel's activity and study its role in cellular communication.

Pathways

CACNA2D1 actively participates in electrical signaling and muscle contraction pathways. It contributes to the excitation-contraction coupling pathway in muscle cells aligning closely with proteins like the ryanodine receptor ensuring efficient calcium-induced calcium release. Its action also extends to the neurotransmitter release pathway where it associates with synaptic proteins regulating neurotransmission. The function within these pathways underlines its importance in both muscle and neural activities.

CACNA2D1 associates with conditions such as epilepsy and neuropathic pain. The dysfunction in its channel activity can lead to altered calcium flow contributing to these disorders. The alpha-2-delta ligand such as the gabapentin chemical structure plays a role here often targeting the channel to mitigate symptoms. Gabapentin's influence highlights the therapeutic potential of targeting CACNA2D1 in disease management providing critical insights into neuromodulation treatments.
style
left-column
style
left-column

Quality control

STR analysis

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

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column

Target data

See full target information CACNA2D1
style
left-column
style
left-column
style
right-column

Complementary Products

Primary Antibodies

AB253190

Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [EPR23267-8]

RabMAb|Recombinant|Trial Size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium channel L type DHPR alpha 2 subunit/CACNA2D1 antibody [EPR23267-8] (AB253190)

  • 0
  • 0
Proteins & Peptides

AB313361

Recombinant Human WFDC2 Protein (His tag)

Mass Spectrometry - Recombinant Human WFDC2 Protein (His tag) (AB313361)

  • 0
  • 0
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
Biochemicals

AB120219

Gabapentin, alpha2delta Ligand

Chemical Structure - Gabapentin, alpha2delta Ligand (AB120219)

  • 0
  • 0

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