Human PXN (Paxillin) knockout A-431 cell lysate (ab261701)
Overview
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Product name
Human PXN (Paxillin) knockout A-431 cell lysate
See all Paxillin kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A431 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 7 bp deletion, 1 bp insertion; Frameshift = 99.7% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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, 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 ab280451 - Human PXN knockout A431 cell lysate 1 x 100µg ab263973 - Human wild-type A-431 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Epidermoid Carcinoma -
Gender
Female
Target
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Function
Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). -
Sequence similarities
Belongs to the paxillin family.
Contains 4 LIM zinc-binding domains. -
Post-translational
modificationsPhosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens. -
Cellular localization
Cytoplasm > cytoskeleton. Cell junction > focal adhesion. - Information by UniProt
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Alternative names
- FLJ16691
- FLJ23042
- Paired box protein Pax 1
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 ab261701 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.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. |
Images
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Western blot - Human PXN (Paxillin) knockout A-431 cell lysate (ab261701)Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 2: PXN knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 3: U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate 20 ug
Lane 4: PC3 (Human prostate adenocarcinoma cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab32115 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32115 was shown to specifically react with PXN in wild-type A-431 cells as signal was lost in PXN knockout cell line ab261892 (knockout cell lysate ab261701). Wild-type and PXN knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. Ab32115 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
Western blot - Human PXN (Paxillin) knockout A-431 cell lysate (ab261701)Lane 1: Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 2: PXN knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug
Lane 3: U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate 20 ug
Lane 4: PC3 (Human prostate adenocarcinoma cell line) whole cell lysate 20 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab23510 observed at 65 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab23510 was shown to recognize PXN in wild-type A-431 cells as signal was lost at the expected MW in PXN knockout cell line ab261892 (knockout cell lysate ab261701). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and PXN knockout samples were subjected to SDS-PAGE. Ab23510 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
Next Generation Sequencing - Human PXN (Paxillin) knockout A-431 cell lysate (ab261701)X = 7 bp deletion, 1 bp insertion
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)
ab261701 has not yet been referenced specifically in any publications.