Human S100A4 knockout HeLa cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human S100A4 knockout HeLa cell line (AB265709)
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 55 kDa.
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 ° 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 (<a href='/en-us/products/cell-lysates/human-s100a4-knockout-hela-cell-lysate-ab257046'>ab257046</a>) at 20 µg
Predicted band size: 12 kDa
Observed band size: 11 kDa
false
- WB
Lab
Western blot - Human S100A4 knockout HeLa cell line (AB265709)
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 55 kDa.
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 ° 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 (<a href='/en-us/products/cell-lysates/human-s100a4-knockout-hela-cell-lysate-ab257046'>ab257046</a>) at 20 µg
Predicted band size: 12 kDa
Observed band size: 11 kDa
false
- WB
Lab
Western blot - Human S100A4 knockout HeLa cell line (AB265709)
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 55 kDa.
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 ° 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 (ab265709)
Predicted band size: 12 kDa
Observed band size: 11 kDa
false
- WB
Lab
Western blot - Human S100A4 knockout HeLa cell line (AB265709)
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 HeLa 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 °. 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 (ab265709)
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
- WB
Lab
Western blot - Human S100A4 knockout HeLa cell line (AB265709)
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 55 kDa.
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 ° 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 (ab265709)
Predicted band size: 12 kDa
Observed band size: 11 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human S100A4 knockout HeLa cell line (AB265709)
Allele-1 : 5 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human S100A4 knockout HeLa cell line (AB265709)
Allele-2 : 1 bp insertion in exon 2.
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)](https://content.abcam.com/products/images/s100a4-antibody-epr146392-ab197896--western-blot-img165500.jpg)
- 4
- 9
Reactivity data
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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