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AB257046

Human S100A4 knockout HeLa cell lysate

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S100A4 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 insertion in exon2 and 5 bp deletion in exon2.
7 Images
Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)
  • WB

Lab

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : S100A4 knockout HeLa cell lysate 20 μg
Lanes 1 - 2 : Merged signal (red and green). Green - ab218511 observed at 11 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab218511 was shown to react with S100A4 in wild-type HeLa cells in Western blot with loss of signal observed in S100A4 knockout cell line ab265709 (S100A4 knockout cell lysate ab257046). Wild-type HeLa and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab218511 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 0.5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-S100A4 antibody [S100A4/1481] (<a href='/en-us/products/primary-antibodies/s100a4-antibody-s100a4-1481-ab218511'>ab218511</a>) at 0.5 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

S100A4 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-s100a4-knockout-hela-cell-line-ab265709'>ab265709</a>)

Predicted band size: 12 kDa

Observed band size: 11 kDa

false

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)
  • WB

Lab

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : S100A4 knockout HeLa cell lysate 20 μg
Lane 3 : Wild-type A549 cell lysate 20 μg
Lane 4 : S100A4 knockout A549 cell lysate 20 μg
False colour image of Western blot : Anti-S100A4 antibody [EPR14639(2)] staining at 1/1000 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab197896 was shown to bind specifically to S100A4. A band was observed at 11 kDa in wild-type and A549 cell lysates with no signal observed at this size in S100A4 knockout HeLa cell line ab265709 (knockout cell lysate ab257046) and S100A4 knockout A549 cell line ab261865 (knockout cell lysate ab261674). To generate this image, wild-type and S100A4 knockout HeLa and S100A4 knockout A549 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-S100A4 antibody [EPR14639(2)] (<a href='/en-us/products/primary-antibodies/s100a4-antibody-epr146392-ab197896'>ab197896</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

S100A4 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-s100a4-knockout-hela-cell-line-ab265709'>ab265709</a>)

Lane 3:

Wild-type A549 cell lysate at 20 µg

Lane 4:

S100A4 knockout A549 cell lysate at 20 µg

Predicted band size: 12 kDa

Observed band size: 11 kDa

false

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)
  • WB

Lab

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : S100A4 knockout HeLa cell lysate 20 μg
Lanes 1 - 2 : Merged signal (red and green). Green - ab218512 observed at 11 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
ab218512 was shown to react with S100A4 in wild-type HeLa cells in Western blot with loss of signal observed in S100A4 knockout cell line ab265709 (S100A4 knockout cell lysate ab257046). Wild-type HeLa and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab218512 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 0.5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-S100A4 antibody [S100A4/1482] (<a href='/en-us/products/primary-antibodies/s100a4-antibody-s100a4-1482-ab218512'>ab218512</a>) at 0.5 µg/mL

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

S100A4 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-s100a4-knockout-hela-cell-line-ab265709'>ab265709</a>)

Predicted band size: 12 kDa

Observed band size: 11 kDa

false

Sanger Sequencing - Human S100A4 knockout HeLa cell lysate (AB257046)
  • Sanger seq

Unknown

Sanger Sequencing - Human S100A4 knockout HeLa cell lysate (AB257046)

Allele-1 : 5 bp deletion in exon2

Sanger Sequencing - Human S100A4 knockout HeLa cell lysate (AB257046)
  • Sanger seq

Unknown

Sanger Sequencing - Human S100A4 knockout HeLa cell lysate (AB257046)

Allele-2 : 1 bp insertion in exon2

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)
  • WB

Lab

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)

Lane 1 : Wild-type HeLa cell lysate 20 μg
Lane 2 : S100A4 knockout HeLa cell lysate 20 μg
Lanes 1 - 2 : Merged signal (red and green). Green - ab124805 observed at 11 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab124805 was shown to react with S100A4 in wild-type HeLa cells in Western blot with loss of signal observed in S100A4 knockout cell line ab265709 (S100A4 knockout cell lysate ab257046). Wild-type HeLa and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab124805 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-S100A4 antibody [EPR2761(2)] (<a href='/en-us/products/primary-antibodies/s100a4-antibody-epr27612-ab124805'>ab124805</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout HeLa cell lysate (ab257046) at 20 µg

Predicted band size: 12 kDa

Observed band size: 11 kDa

false

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)
  • WB

Lab

Western blot - Human S100A4 knockout HeLa cell lysate (AB257046)

Lane 1 : Wild-type HeLa cell lysate μg
Lane 2 : S100A4 knockout HeLa cell lysate 20 μg
Lanes 1 - 2 : Merged signal (red and green). Green - ab197896 observed at 11 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab197896 was shown to react with S100A4 in wild-type HeLa cells in Western blot with loss of signal observed in S100A4 knockout cell line ab265709 (S100A4 knockout cell lysate ab257046). Wild-type HeLa and S100A4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab197896 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-S100A4 antibody [EPR14639(2)] (<a href='/en-us/products/primary-antibodies/s100a4-antibody-epr146392-ab197896'>ab197896</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout HeLa cell lysate (ab257046) at 20 µg

Predicted band size: 12 kDa

Observed band size: 11 kDa

false

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 insertion in exon2 and 5 bp deletion in exon2.

Disease

Adenocarcinoma

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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":"ab257046-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human S100A4 knockout HeLa cell lysate", "number":"AB257046-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HeLa cell lysate", "number":"AB257046-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
S100A4
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.

S100A4 also known as metastasin or MTS1 is a protein with a molecular weight of approximately 12 kDa. It functions by binding calcium which induces conformational changes aiding in its interaction with target proteins. S100A4 is part of the S100 family of proteins characterized by two EF-hand calcium-binding motifs. Expression of S100A4 is observed in various cell types including fibroblasts endothelial cells and several tumor cells where it contributes to processes like cell motility invasion and angiogenesis.
Biological function summary

The role of S100A4 extends beyond mere calcium binding; it associates with the cytoskeleton promoting cell structures related to movement and metastasis. It does not act alone but often partners with proteins like myosin IIA to regulate cytoskeletal dynamics. Additionally S100A4 modulates extracellular matrix components and interacts with signaling pathways that control cancer progression making it significant in both normal physiological contexts and pathological states.

Pathways

Functional activities of S100A4 integrate into important pathways involved in cellular movement and metastasis. S100A4 plays roles in the regulation of the Wnt/β-catenin pathway which is pivotal for cell-cell interaction and fate determination. It also interacts with matrix metalloproteinases like MMPs thereby managing processes of tissue invasion and remodeling essential in cancer metastasis. Other pathways influencing cell adhesion and signal transduction may indirectly involve S100A4 through related proteins.

Elevated S100A4 levels have implications in cancer particularly in enhancing metastasis in breast and colorectal cancers. The protein promotes tumor developments by inducing changes in the tumor microenvironment making it a target of interest in oncology. S100A4 also interacts with proteins like p53 and β-catenin which are critical in the pathology of tumors. Moreover S100A4 contributes to fibrotic diseases accentuating tissue remodeling activities by modulating fibroblast behavior and extracellular matrix deposition.
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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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