Human SIPA1 (Spa-1) knockout HeLa cell lysate
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SIPA1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 182 bp deletion in exon5 and 1 bp deletion in exon5 and 4 bp deletion in exon5.
View Alternative Names
GTPase-activating protein Spa-1, MGC102688, MGC17037, SIPA1_HUMAN, SP A1, Signal induced proliferation associated 1, Signal induced proliferation associated gene 1, Signal-induced proliferation-associated protein 1, p130 SPA-1
- WB
Lab
Western blot - Human SIPA1 (Spa-1) knockout HeLa cell lysate (AB258189)
Lane 1 : Wild-type HeLa cell lysate (20 μg)
Lane 2 : SIPA1 knockout HeLa cell lysate (20 μg)
Lane 3 : Raji cell lysate (20 μg)
Lane 4 : HEK-293 cell lysate (20 μg)
Lanes 1-4 : Merged signal (red and green). Green - ab189929 observed at 130 kDa. Red - loading control ab7291 observed at 50 kDa.
ab189929 Anti-Spa-1 antibody [EPR14134] was shown to specifically react with Spa-1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265434 (knockout cell lysate ab258189) was used. Wild-type and Spa-1 knockout samples were subjected to SDS-PAGE. ab189929 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 5000 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.
All lanes:
Western blot - Anti-Spa-1 antibody [EPR14134] (<a href='/en-us/products/primary-antibodies/spa-1-antibody-epr14134-ab189929'>ab189929</a>) at 1/5000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Spa-1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human SIPA1 (Spa-1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-sipa1-spa-1-knockout-hela-cell-line-ab265434'>ab265434</a>)
Lane 3:
Raji cell lysate at 20 µg
Lane 4:
HEK-293 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Predicted band size: 112 kDa
Observed band size: 130 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SIPA1 (Spa-1) knockout HeLa cell lysate (AB258189)
Allele-2 : 1 bp deletion in exon5
- Sanger seq
Unknown
Sanger Sequencing - Human SIPA1 (Spa-1) knockout HeLa cell lysate (AB258189)
Allele-3 : 182 bp deletion in exon5
- Sanger seq
Unknown
Sanger Sequencing - Human SIPA1 (Spa-1) knockout HeLa cell lysate (AB258189)
Allele-1 : 4 bp deletion in exon5
Reactivity data
Product details
Knockout cell lysate achieved by CRISPR/Cas9.
REACH authorisation
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.
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. 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.
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.
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
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Spa-1 engages in the control of cell adhesion and migration through its interaction with the Ras-related protein Rap1. As part of larger signal transduction complexes Spa-1 influences the organization of actin cytoskeleton and impacts integrin-mediated cell adhesion. Through its GAP activity it maintains cellular responsiveness to external stimuli impacting processes like immune cell trafficking and neuronal growth. Studies have noted its significance in maintaining homeostasis in tissues where it is expressed.
Pathways
Spa-1 integrates into key signaling cascades impacting the Ras superfamily of GTPases. Within the Rap1 signaling pathway Spa-1 works alongside proteins like RapGEF to finely regulate signal transduction processes affecting cell adhesion and cytoskeletal dynamics. In the context of the integrin signaling pathway Spa-1 modulates the actions of B-Raf in connection with Rap1 dictating responses necessary for cellular motility and positioning.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Western blot - Anti-Spa-1 antibody [EPR14134] (AB189929)](https://content.abcam.com/products/images/spa-1-antibody-epr14134-ab189929--western-blot-img106572.png)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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