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AB255379

Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line

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COL4A1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 25. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 이미지
Western blot - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (AB255379)
  • WB

Lab

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] (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='/ko/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 (ab255379)

Predicted band size: 161 kDa

Observed band size: 200 kDa

false

Sanger Sequencing - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (AB255379)
  • Sanger seq

Unknown

Sanger Sequencing - Human COL4A1 (Collagen IV alpha 1) knockout HeLa cell line (AB255379)

Homozygous : 1 bp insertion in exon 25.

주요 정보

세포 유형

HeLa

Species or organism

Human

조직

Cervix

제형

Liquid

form

Knockout 검증

Sanger Sequencing

Mutation 정보

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 25

질병

Adenocarcinoma

Reactivity 정보

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

제품 세부 정보

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.

제품 구성

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특성 및 보관 정보

유전자명
COL4A1
유전자 편집 종류
Knockout
유전자 편집 방식
CRISPR technology
Knockout 검증
Sanger Sequencing
Zygosity
Homozygous
배송 시 보관 조건
Dry Ice
적절한 단기 보관 조건
-196°C
적절한 장기 보관 조건
-196°C

추가 정보

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

품질 관리

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

초기 취급 지침

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

Subculture 가이드라인
  • 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.
배양 배지

DMEM (High Glucose) + 10% FBS

동결보존 배지

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

제품 프로토콜

Product promise

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