COL4A1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 25.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 25
Alternative names
Arresten, COL4A1 NC1 domain, Collagen IV alpha-1 polypeptide, Collagen alpha-1(IV) chain, Collagen of basement membrane, alpha 1 chain, HANAC, POREN1
Recommended products
COL4A1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 25.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 25
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- COL4A1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Collagen IV alpha 1 also known as COL4A1 forms an essential component of type IV collagen a major structural protein in basement membranes. With a molecular mass of approximately 185 kDa it contributes significantly to the formation of the structural matrix that supports epithelial and endothelial cells. Expression of COL4A1 is widespread notably in tissues with high involvement of basement membranes such as kidneys the eye and the cerebral vasculature. It takes part in the assembly of the collagen IV network essential for a stable extracellular environment.
- 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.
- Associated diseases and disorders
Mutations or abnormalities in collagen type IV alpha 1 relate to conditions like cerebral small vessel disease (SVD) and Alport syndrome. SVD is associated with compromised vascular integrity where COL4A1 mutations may disrupt normal blood vessel function. In Alport syndrome disruptions in type IV collagen may lead to progressive kidney disease due to the protein’s vital role in glomerular basement membrane structure. Furthermore COL4A1’s interaction with proteins like COL4A5 highlights its broader impact in these disorders as COL4A5 mutations are a known cause of Alport syndrome.
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2 product images
Western blot - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (ab255379)
False colour image of Western blot: Anti-Collagen IV antibody [EPR20966] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Collagen IV antibody [EPR20966] 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 Human COL4A1 (Collagen IV alpha 1) knockout HeLa 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-Collagen IV antibody [EPR20966] (Anti-Collagen IV antibody [EPR20966] ab214417) 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 (ab255379)
Performed under reducing conditions.
Predicted band size: 161 kDa
Observed band size: 200 kDa
Sanger Sequencing - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (ab255379)
Homozygous: 1 bp insertion in exon 25.
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