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AB261865

Human S100A4 knockout A549 cell line

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S100A4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 95%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
6 Images
Western blot - Human S100A4 knockout A549 cell line (AB261865)
  • WB

Lab

Western blot - Human S100A4 knockout A549 cell line (AB261865)

Lanes 1 - 4 : Merged signal (red and green). Green - ab124805 observed at 12 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab124805 was shown to react with S100A4 in wild-type A549 cells in Western blot with loss of signal observed in S100A4 knockout cell line ab261865 (knockout cell lysate ab261674). Wild-type A549 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 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 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 A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout A549 cell line (ab261865)

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

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Western blot - Human S100A4 knockout A549 cell line (AB261865)
  • WB

Lab

Western blot - Human S100A4 knockout A549 cell line (AB261865)

Lanes 1 - 4 : Merged signal (red and green). Green - ab218512 observed at 12 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab218512 was shown to specifically react with S100A4 in wild-type A549 cells as signal was lost in S100A4 knockout cell line ab261865 (knockout cell lysate ab261674). Wild-type and S100A4 knockout samples were subjected to SDS-PAGE. ab218512 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour 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 1 µg/mL

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Human S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout A549 cell line (ab261865)

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

Western blot - Human S100A4 knockout A549 cell line (AB261865)
  • WB

Lab

Western blot - Human S100A4 knockout A549 cell line (AB261865)

Lanes 1 - 4 : Merged signal (red and green). Green - ab218511 observed at 12 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab218511 was shown to specifically react with S100A4 in wild-type A549 cells as signal was lost in S100A4 knockout cell line ab261865 (knockout cell lysate ab261674). Wild-type and S100A4 knockout samples were subjected to SDS-PAGE. ab218511 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour 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 1 µg/mL

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout A549 cell line (ab261865)

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 12 kDa

Observed band size: 12 kDa

false

Western blot - Human S100A4 knockout A549 cell line (AB261865)
  • WB

Lab

Western blot - Human S100A4 knockout A549 cell line (AB261865)

Lanes 1 - 4 : Merged signal (red and green). Green - ab41532 observed at 12 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab41532 was shown to recognize S100A4 in wild-type A549 cells as signal was lost at the expected MW in S100A4 knockout cell line ab261865 (knockout cell lysate ab261674). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and S100A4 knockout samples were subjected to SDS-PAGE. ab41532 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Anti-S100A4 antibody (<a href='/en-us/products/unavailable/s100a4-antibody-ab41532'>ab41532</a>) at 1/250 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human S100A4 knockout A549 cell line (ab261865)

Lane 3:

A-375 (Human malignant melanoma cell line) whole cell lysate, 20 ug at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

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Next Generation Sequencing - Human S100A4 knockout A549 cell line (AB261865)
  • NGS

Supplier Data

Next Generation Sequencing - Human S100A4 knockout A549 cell line (AB261865)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 95%

Next Generation Sequencing - Human S100A4 knockout A549 cell line (AB261865)
  • NGS

Supplier Data

Next Generation Sequencing - Human S100A4 knockout A549 cell line (AB261865)

1 bp insertion after His16 of the WT protein

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 95%

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB197896

Anti-S100A4 antibody [EPR14639(2)]

RabMAb|Recombinant|KO Validated|20ul selling size

Western blot - Anti-S100A4 antibody [EPR14639(2)] (AB197896)

  • 4
  • 8
Assay Kits

AB283547

Human S100A4 ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human S100A4 ELISA Kit (AB283547)

  • 0
  • 0
Cell Lines & Lysates

AB266658

Human CD276 knockout HEK-293T cell line

Advanced Validation

Western blot - Human CD276 knockout HEK-293T cell line (AB266658)

  • 5
  • 2
Proteins & Peptides

AB109341

Recombinant Mouse S100A4 protein

SDS-PAGE - Recombinant Mouse S100A4 protein (AB109341)

  • 0
  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "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

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.

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What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab261865-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab259777 Human wild-type A549 cell line", "number":"AB261865-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab261865 Human S100A4 knockout A549 cell line", "number":"AB261865-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab261865-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab261865 Human S100A4 knockout A549 cell line", "number":"AB261865-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
S100A4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

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

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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

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information S100A4
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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'.

For licensing inquiries, please contact partnerships@abcam.com