Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell lysate
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- WB
Lab
Western blot - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell lysate (AB263755)
Lane 1 : Wild-type HeLa cell lysate 20 ug
Lane 2 : Col4a1 knockout HeLa cell lysate 20 ug
False colour image of Western blot : Anti-Collagen IV antibody [EPR20966] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab214417 was shown to bind specifically to Collagen IV. A band was observed at 200 kDa in wild-type HeLa cell lysates with no signal observed at this size in Col4a1 knockout cell line ab255379 (knockout cell lysate ab263755). To generate this image, wild-type and Col4a1 knockout HeLa 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Collagen IV antibody [EPR20966] (<a href='/en-us/products/primary-antibodies/collagen-iv-antibody-epr20966-ab214417'>ab214417</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
Col4a1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-col4a1-collagen-iv-alpha-1-knockout-hela-cell-line-ab255379'>ab255379</a>)
Predicted band size: 161 kDa
Observed band size: 200 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell lysate (AB263755)
Homozygous : 1 bp insertion in exon 25
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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, 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
Zygosity
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
Collagen type IV alpha 1 contributes to the structural integrity and mechanical stability of the basement membrane. It operates as part of a heterotrimeric complex alongside COL4A2 integrating into the basement membrane’s unique mesh-like structure. This arrangement is important for the maintenance of cell attachment movement and the filtration barrier in tissues like the glomeruli. This protein also facilitates separations between tissue compartments helping organize cell layers into functional units.
Pathways
The collagen type IV alpha 1 protein participates in the integrin signaling pathway and the matrix metalloproteinase (MMP) pathway. It interacts with other collagen forms and proteins such as integrins involved in signaling events that regulate cell survival proliferation and migration. Its involvement in the MMP pathway highlights its role in tissue remodeling and repair as MMPs modulate the extracellular environment including basement membrane degradation and reconstruction.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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