Human PON2 knockout HeLa cell lysate (ab258128)
Overview
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Product name
Human PON2 knockout HeLa cell lysate
See all PON2 kits -
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 exon4. -
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.
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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 ab261149 - Human PON2 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
Capable of hydrolyzing lactones and a number of aromatic carboxylic acid esters. Has antioxidant activity. Is not associated with high density lipoprotein. Prevents LDL lipid peroxidation, reverses the oxidation of mildly oxidized LDL, and inhibits the ability of MM-LDL to induce monocyte chemotaxis. -
Tissue specificity
Widely expressed with highest expression in liver, lung, placenta, testis and heart. -
Sequence similarities
Belongs to the paraoxonase family. -
Post-translational
modificationsThe signal sequence is not cleaved. -
Cellular localization
Membrane. - Information by UniProt
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Alternative names
- A esterase 2
- A-esterase 2
- Aromatic esterase 2
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab258128 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 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 39 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 39 kDa. |
Images
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Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: PICALM knockout HeLa cell lysate (20 µg)
Lane 3: PON2 cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab183710 observed at 39,41 kDa. Red - loading control ab7291 observed at 50 kDa.
ab183710 Anti-PON2 antibody [EPR15295-82] was shown to specifically react with PON2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265927 (knockout cell lysate ab258128) was used. Wild-type and PON2 knockout samples were subjected to SDS-PAGE. ab183710 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C 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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Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: PICALM knockout HeLa cell lysate (20 µg)
Lane 3: PON2 cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab40969 observed at 39,41 kDa. Red - loading control ab7291 observed at 50 kDa.
ab40969 Anti-PON2 antibody was shown to specifically react with PON2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265927 (knockout cell lysate ab258128) was used. Wild-type and PON2 knockout samples were subjected to SDS-PAGE. ab40969 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C 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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Homozygous: 1 bp deletion in exon4
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)
ab258128 has not yet been referenced specifically in any publications.