AB265748

Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line

Advanced Validation
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PRKAR2A 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 1 and 4 bp deletion in exon 1.

View Alternative Names

KAP2_HUMAN, MGC3606, PRKA R2, PRKAR 2, PRKAR2, PRKAR2A, Protein kinase A RII alpha subunit, Protein kinase cAMP dependent regulatory type II alpha, cAMP dependent protein kinase regulatory subunit RII alpha, cAMP dependent protein kinase regulatory subunit alpha 2, cAMP dependent protein kinase type II alpha regulatory chain, cAMP-dependent protein kinase type II-alpha regulatory subunit

3 Images

Lab

Western blot - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Lanes 1-3 : Merged signal (red and green). Green - ab32514 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

Unpurified ab32514 Anti-PKA R2/PKR2 antibody [Y116] was shown to specifically react with PKA R2/PKR2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265748 (knockout cell lysate ab257607) was used. Wild-type and PKA R2/PKR2 knockout samples were subjected to SDS-PAGE. ab32514 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 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-PKA R2/PKR2 antibody [Y116] (<a href='/en-us/products/primary-antibodies/pka-r2-pkr2-antibody-y116-ab32514'>ab32514</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PRKAR2A knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (ab265748)

Lane 3:

K-562 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 50 kDa

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Sanger Sequencing - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Sanger seq

Unknown

Sanger Sequencing - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Allele-1 : 4 bp deletion in exon 1.

Sanger seq

Unknown

Sanger Sequencing - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Allele-2 : 1 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, 1 bp deletion in exon 1 and 4 bp deletion in exon 1

Antibiotic resistance

Puromycin 1µg/mL

Disease

Adenocarcinoma

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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

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 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

PRKAR2A

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.

Protein Kinase A R2 (PKR2) also known as PRKAR2A is a regulatory subunit of Protein Kinase A (PKA). PKA is part of a larger complex known as the cAMP-dependent protein kinase which plays an important role in cellular signaling. PKA consists of regulatory and catalytic subunits with PKR2 serving as one of its regulatory forms. PKR2 possesses a molecular mass of approximately 47 kDa and is available in various human tissues with notable expression in the brain and cardiovascular system.

Biological function summary

PKR2 regulates the activity of PKA by binding to the catalytic subunit in the absence of cAMP therefore inhibiting its activity. When cAMP is present it binds to the regulatory subunit PKR2 leading to the release and activation of the catalytic subunit of PKA. PKR2 is an integral part of the PKA holoenzyme complex which plays a critical role in mediating cellular responses to hormonal stimulation.

Pathways

PKR2 is heavily involved in the cAMP-dependent signaling pathway. This pathway is essential for translating extracellular signals into cellular actions. It interacts with various effector proteins including adenylate cyclases and phosphodiesterases to modulate responses such as gene transcription metabolism and memory formation. PKR2-related activity influences pathways shared with proteins like the G-protein subunits which facilitate the effects of neurotransmitters.

PKR2 irregularities can impact conditions like heart failure and neurodegenerative diseases. Disrupted PKA pathway signaling involving PKR2 can contribute to cardiac dysfunction by affecting heart muscle contractility and hypertrophy. In neurodegenerative diseases the improper functioning of PKR2 can exacerbate disruptions in neural signaling. A connection exists between PKR2 and proteins such as CREB (cAMP response element-binding protein) which links PKR2's role to memory deficits observed in neurodegeneration.

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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

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information PRKAR2A

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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

Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line

Western blot - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Sanger Sequencing - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Sanger Sequencing - Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell line (AB265748)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Antibiotic resistance

Disease

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Handling procedures

Initial handling guidelines

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Target data

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Proteins & Peptides

Product promise