Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell lysate (ab257607)
Overview
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Product name
Human PRKAR2A (PKA R2/PKR2) knockout HeLa cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 4 bp deletion in exon 1. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.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.
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab261021 - Human PRKAR2A knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8, 12 CSF1PO: 9, 10
Target
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Function
Regulatory subunit of the cAMP-dependent protein kinases involved in cAMP signaling in cells. Type II regulatory chains mediate membrane association by binding to anchoring proteins, including the MAP2 kinase. -
Tissue specificity
Four types of regulatory chains are found: I-alpha, I-beta, II-alpha, and II-beta. Their expression varies among tissues and is in some cases constitutive and in others inducible. -
Sequence similarities
Belongs to the cAMP-dependent kinase regulatory chain family.
Contains 2 cyclic nucleotide-binding domains. -
Post-translational
modificationsPhosphorylated by the activated catalytic chain. -
Cellular localization
Cytoplasm. Cell membrane. Colocalizes with PJA2 in the cytoplasm and the cell membrane. - Information by UniProt
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Alternative names
- cAMP dependent protein kinase regulatory subunit alpha 2
- cAMP dependent protein kinase regulatory subunit RII alpha
- cAMP dependent protein kinase type II alpha regulatory chain
see all
Associated products
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KO cell lines
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Related Products
Images
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Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PRKAR2A knockout HeLa cell lysate (20 ug)
Lane 3:K-562 cell lysate (20 ug)ab32514 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.
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Allele-1: 4 bp deletion in exon 1
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Allele-2: 1 bp deletion in exon 1
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab257607 has not yet been referenced specifically in any publications.