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AB257406

Human CYR61 (CCN1) knockout HeLa cell lysate

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CCN1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and Insertion of the selection cassette in exon1.
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Western blot - Human CYR61 (CCN1) knockout HeLa cell lysate (AB257406)
  • WB

Lab

Western blot - Human CYR61 (CCN1) knockout HeLa cell lysate (AB257406)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : CYR61 knockout HeLa cell lysate (20 ug)
Lane 3 : MDA-MB-231 cell lysate (20 ug)
Lane 4 : MCF7 cell lysate (20 ug)

ab230947 was shown to specifically react with CYR61/CCN1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265288 (knockout cell lysate ab257406) was used. Wild-type and CYR61/CCN1 knockout samples were subjected to SDS-PAGE. ab230947 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC at 1 in 500 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-CYR61/CCN1 antibody [EPR20681] (<a href='/en-us/products/primary-antibodies/cyr61-ccn1-antibody-epr20681-ab230947'>ab230947</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CYR61 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CYR61 (CCN1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cyr61-ccn1-knockout-hela-cell-line-ab265288'>ab265288</a>)

Lane 3:

MDA-MB-231 cell lysate at 20 µg

Lane 4:

MCF7 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 1/10000 dilution

Predicted band size: 42 kDa

Observed band size: 47 kDa

false

Sanger Sequencing - Human CYR61 (CCN1) knockout HeLa cell lysate (AB257406)
  • Sanger seq

Unknown

Sanger Sequencing - Human CYR61 (CCN1) knockout HeLa cell lysate (AB257406)

Allele-1 : 1 bp deletion in exon1

Sanger Sequencing - Human CYR61 (CCN1) knockout HeLa cell lysate (AB257406)
  • Sanger seq

Unknown

Sanger Sequencing - Human CYR61 (CCN1) knockout HeLa cell lysate (AB257406)

Allele-2 : Insertion of the selection cassette in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and Insertion of the selection cassette in exon1.

Disease

Adenocarcinoma

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Reactivity data

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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.

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What's included?

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Properties and storage information

Gene name
CCN1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

The CYR61/CCN1 protein also known simply as CYR61 CCN1 or CYR61 protein is a cellular protein with a molecular weight of approximately 40 kDa. It is a member of the CCN family of regulatory proteins which includes connective tissue growth factors. CYR61 is secreted into the extracellular matrix and has a critical role in cell adhesion and signaling. The protein is expressed in various tissues with higher levels observed in the placenta lungs and kidneys reflecting its role in organ development and repair processes.
Biological function summary

The CYR61/CCN1 protein acts as a signaling molecule that influences cell proliferation migration and adhesion. It interacts with integrins and heparan sulfate proteoglycans on the cell surface facilitating cellular communication that is essential for structural and functional tissue integrity. Additionally CYR61 is a part of multiprotein complexes impacting angiogenesis and inflammation. These activities place CYR61/CCN1 at a pivotal point in wound healing and vascularization processes.

Pathways

The CYR61/CCN1 protein participates in key signaling pathways such as the MAPK/ERK and Wnt pathways. These pathways modulate gene expression related to cell growth and differentiation. CYR61 collaborates with other proteins including VEGF and FGF to regulate cardiovascular development and morphogenesis. The interaction with these pathways highlights the protein’s integral role in cellular homeostasis and response to physiological changes.

CYR61/CCN1 is linked to cancer progression and fibrotic diseases. Elevated levels of this protein are frequently found in breast cancer emphasizing its role in tumor growth and metastasis. It associates with other proteins like TGF-beta during fibrotic responses contributing to the pathology of fibrosis in organs such as the liver and lungs. The involvement in these processes highlights its potential as a therapeutic target in oncology and fibrotic disease management.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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