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AB256957

Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate

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ITGA6 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 exon2 and 1 bp insertion in exon2.
4 Images
Western blot - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)
  • WB

Lab

Western blot - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)

Lane 1 : Wild-type SW480 cell lysate, 20 ug

Lane 2 : ITGA6 knockout SW480 cell lysate, 20 ug

Lane 3 : HEK-293 cell lysate, 20 ug

Lanes 1 - 3 : Merged signal (red and green). Green - ab124924 observed at 130 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab124924 was shown to react with Integrin alpha 6 in wild-type SW480 cells in western blot with loss of signal observed in ITGA6 knockout cell line ab265838 (knockout cell lysate ab256957). Wild-type and ITGA6 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab124924 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin alpha 6 antibody [EPR5578] (<a href='/en-us/products/primary-antibodies/integrin-alpha-6-antibody-epr5578-ab124924'>ab124924</a>) at 1/1000 dilution

Lane 1:

Wild-type SW480 cell lysate at 20 µg

Lane 2:

ITGA6 knockout SW480 cell lysate at 20 µg

Lane 3:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 126 kDa

Observed band size: 130 kDa

false

Western blot - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)
  • WB

Lab

Western blot - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)

Lane 1 : Wild-type SW480 cell lysate, 20 ug

Lane 2 : ITGA6 knockout SW480 cell lysate, 20 ug

Lane 3 : HEK-293 cell lysate, 20 ug

Lanes 1 - 3 : Merged signal (red and green). Green - ab181551 observed at 130 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab181551 was shown to react with Integrin alpha 6 in wild-type SW480 cells in western blot with loss of signal observed in ITGA6 knockout cell line ab265838 (knockout cell lysate ab256957). Wild-type and ITGA6 knockout SW480 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab181551 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Integrin alpha 6 antibody [EPR18124] (<a href='/en-us/products/primary-antibodies/integrin-alpha-6-antibody-epr18124-ab181551'>ab181551</a>) at 1/2000 dilution

Lane 1:

Wild-type SW480 cell lysate at 20 µg

Lane 2:

ITGA6 knockout SW480 cell lysate at 20 µg

Lane 3:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat anti-Mouse IgG H&L preabsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 126 kDa

Observed band size: 130 kDa

false

Sanger Sequencing - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)
  • Sanger seq

Unknown

Sanger Sequencing - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)

Allele-2 : 1 bp insertion in exon2

Sanger Sequencing - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)
  • Sanger seq

Unknown

Sanger Sequencing - Human ITGA6 (Integrin alpha 6) knockout HeLa cell lysate (AB256957)

Allele-1 : 1 bp deletion in exon2

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 exon2 and 1 bp insertion in exon2.

Disease

Adenocarcinoma

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Primary Antibodies

AB181551

Anti-Integrin alpha 6 antibody [EPR18124]

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 6 antibody [EPR18124] (AB181551)

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Recombinant human Vitronectin/S-Protein (Animal Free)

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Reactivity data

{ "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>" } } }
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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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab256957-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ITGA6 knockout HeLa cell lysate", "number":"AB256957-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB256957-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
ITGA6
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.

Integrin alpha 6 also known as ITGA6 or CD49f functions as a transmembrane receptor protein involved in cell adhesion and signal transduction. As part of the broader integrin family integrin alpha 6 typically pairs with the beta 1 or beta 4 subunits forming the alpha6beta1 or alpha6beta4 integrins respectively. It has a molecular mass of approximately 150 kDa. This protein is mainly expressed in epithelial cells but also found in platelets certain immune cells and in the nervous system. Its expression patterns highlight its role in a variety of cellular and tissue functions.
Biological function summary

The integrin alpha 6 protein partners with the beta 4 subunit to form a heterodimer complex that plays an important role in cellular adhesion to the extracellular matrix specifically interacting with laminins. Its main action helps cells attach to the basement membrane which is an important component of epithelial tissue structure and integrity. The alpha 6 integrin provides structural cell support and participates in transmitting signals that influence cell behavior such as movement survival and proliferation.

Pathways

Integrins including the alpha 6 variant are important components in the integrin signaling pathway and are involved in cell motility and survival pathways. In particular the alpha 6 integrin partners with proteins like focal adhesion kinase (FAK) and phosphoinositide 3-kinase (PI3K) to initiate downstream signaling cascades. The interaction with laminins activates pathways critical for developmental processes and tissue remodeling highlighting their role in maintaining cellular environments.

Integrin alpha 6 has significant implications in cancer progression and skin blistering disorders. In various cancers such as breast and skin cancers upregulation of integrin alpha 6 promotes tumor growth and metastasis through interaction with proteins like collagen and laminins found in the tumor microenvironment. Furthermore in the context of blistering disorders such as epidermolysis bullosa mutations in the beta 4 subunit that pairs with alpha 6 can disrupt the integrity of epithelial tissues leading to severe impact on the skin’s structural stability.
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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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