Human PDE4B knockout HeLa cell lysate (ab257576)
Overview
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Product name
Human PDE4B 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, 19 bp deletion in exon12 and 1 bp insertion in exon12. -
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: After reconstitution, store the lysate at -80°C.
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, 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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260288 - Human PDE4B knockout HeLa cell lysate (Lyophilized) 1 x 100µg ab255929 - Human Wild Type HeLa cell lysate (Lyophilized) 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
Hydrolyzes the second messenger cAMP, which is a key regulator of many important physiological processes. May be involved in mediating central nervous system effects of therapeutic agents ranging from antidepressants to antiasthmatic and anti-inflammatory agents. -
Tissue specificity
Expressed in brain, heart, lung and skeletal muscle. -
Pathway
Purine metabolism; 3',5'-cyclic AMP degradation; AMP from 3',5'-cyclic AMP: step 1/1. -
Sequence similarities
Belongs to the cyclic nucleotide phosphodiesterase family. PDE4 subfamily. - Information by UniProt
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Alternative names
- 5''-cyclic phosphodiesterase 4B
- cAMP-specific 3''
- cAMP-specific 3',5'-cyclic phosphodiesterase 4B
see all
Associated products
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KO cell lines
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Related Products
Applications
Our Abpromise guarantee covers the use of ab257576 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | Use at an assay dependent concentration. Predicted molecular weight: 83 kDa. |
Images
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Western blot - Human PDE4B knockout HeLa cell lysate (ab257576)
Lane 1:Wild-type HeLa cell lysate (20 ug)
Lane 2:PDE4B knockout HeLa cell lysate (20 ug)
Lane 3:A549 cell lysate (20 ug)
Lane 4:Mouse brain tissue lysate (20 ug)ab170939 was shown to specifically react with PDE4B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265259 (knockout cell lysate ab257576) was used. Wild-type and PDE4B knockout samples were subjected to SDS-PAGE. ab170939 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 500 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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Sanger Sequencing - Human PDE4B knockout HeLa cell lysate (ab257576)Allele-1: 19 bp deletion in exon12
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Sanger Sequencing - Human PDE4B knockout HeLa cell lysate (ab257576)Allele-2: 1 bp insertion in exon12
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.
References (0)
ab257576 has not yet been referenced specifically in any publications.
