Human PXN (Paxillin) knockout A-431 cell line
- Advanced Validation
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PXN KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 7 bp deletion, 1 bp insertion; Frameshift = 99.7%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
FLJ16691, FLJ23042, PAXI_HUMAN, PXN, PXN protein, Paired box protein Pax 1, Paxillin, Paxillin alpha
- WB
Lab
Western blot - Human PXN (Paxillin) knockout A-431 cell line (AB261892)
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.
All lanes:
Western blot - Anti-Paxillin antibody [M107] (<a href='/en-us/products/primary-antibodies/paxillin-antibody-m107-ab23510'>ab23510</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
PXN knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PXN (Paxillin) knockout A-431 cell line (ab261892)
Lane 3:
U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 20 µg
Lane 4:
PC3 (Human prostate adenocarcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 65 kDa
false
- WB
Lab
Western blot - Human PXN (Paxillin) knockout A-431 cell line (AB261892)
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.
All lanes:
Western blot - Anti-Paxillin antibody [E228] (<a href='/en-us/products/primary-antibodies/paxillin-antibody-e228-ab32115'>ab32115</a>) at 1/10000 dilution
Lane 1:
Wild-type A431 whole cell lysate at 20 µg
Lane 2:
PXN knockout A431 whole cell lysate at 20 µg
Lane 2:
Western blot - Human PXN (Paxillin) knockout A-431 cell line (ab261892)
Lane 3:
U-87 MG whole cell lysate at 20 µg
Lane 4:
PC-3 whole cell lysate at 20 µg
Predicted band size: 65 kDa
Observed band size: 65 kDa
false
Exposure time: 10s
- WB
Lab
Western blot - Human PXN (Paxillin) knockout A-431 cell line (AB261892)
Anti-PXN antibody [Y113] (ab32084) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32084 was shown to bind specifically to PXN. A band was observed at 70 kDa in wild-type A431 cell lysates with no signal observed at this size in PXN knockout cell line ab261892 (knockout cell lysate ab261701). To generate this image, wild-type and PXN knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween$®$ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
All lanes:
Western blot - Anti-Paxillin antibody [Y113] (<a href='/en-us/products/primary-antibodies/paxillin-antibody-y113-ab32084'>ab32084</a>) at 1/1000 dilution
Lane 2:
Western blot - Human PXN (Paxillin) knockout A-431 cell lysate (<a href='/en-us/products/cell-lysates/human-pxn-paxillin-knockout-a-431-cell-lysate-ab261701'>ab261701</a>)
Lane 2:
Western blot - Human PXN (Paxillin) knockout A-431 cell line (ab261892)
Predicted band size: 65 kDa
Observed band size: 70 kDa
false
- WB
Unknown
Western blot - Human PXN (Paxillin) knockout A-431 cell line (AB261892)
Remaining relative intensity is presented.
Loading control : Anti-PPIB
Lane 1:
HEK-293 cells transfected with control siRNA
Lane 2:
HEK-293 cells transfected with target specific siRNA probe
false
- NGS
Supplier Data
Next Generation Sequencing - Human PXN (Paxillin) knockout A-431 cell line (AB261892)
1 bp insertion after Thr135 of the WT protein
- NGS
Lab
Next Generation Sequencing - Human PXN (Paxillin) knockout A-431 cell line (AB261892)
X = 7 bp deletion, 1 bp insertion
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
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
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Paxillin serves key functions in cellular processes such as migration proliferation and survival. It interacts with different proteins including vinculin and talin forming part of a complex at the focal adhesion sites. The phosphorylation state of Paxillin can modulate its interactions and in this way influence the assembly of signaling complexes that control dynamic cellular processes.
Pathways
Paxillin participates in the integrin signaling and MAPK pathways. It operates by linking integrin receptors to the actin cytoskeleton facilitating signal transduction. Paxillin phosphorylation is an important regulatory mechanism within these pathways. In particular its interaction with focal adhesion kinase (FAK) and Src family kinases signifies its role in transmitting signals from the extracellular matrix to the cellular interior which impacts cell behavior and response.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
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Primary Antibodies
AB32084
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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