Human S100A4 knockout HeLa cell lysate (ab257046)
Overview
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Product name
Human S100A4 knockout HeLa cell lysate
See all S100A4 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon2 and 5 bp deletion in exon2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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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Tested applications
Suitable for: Sanger Sequencing, WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260907 - Human S100A4 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Adenocarcinoma -
Gender
Female -
STR Analysis
Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
Target
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Tissue specificity
Ubiquitously expressed. -
Sequence similarities
Belongs to the S-100 family.
Contains 2 EF-hand domains. - Information by UniProt
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Alternative names
- 18A2
- 42A
- calcium Placental protein
see all
Associated products
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Related Products
- Anti-S100A4 antibody [EPR2761(2)] (ab124805)
- Anti-S100A4 antibody [EPR14639(2)] (ab197896)
- Anti-S100A4 antibody [EPR2761(2)] - BSA and Azide free (ab216003)
- Anti-S100A4 antibody [S100A4/1481] (ab218511)
- Anti-S100A4 antibody [EPR14639(2)] - BSA and Azide free (ab220213)
- Anti-S100A4 antibody [EPR2761(2)] - BSA and Azide free (Capture) (ab281227)
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab257046 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sanger Sequencing |
Use at an assay dependent concentration.
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| WB |
Use at an assay dependent concentration. Predicted molecular weight: 12 kDa.
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| Notes |
|---|
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Sanger Sequencing
Use at an assay dependent concentration. |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 12 kDa. |
Images
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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. -
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. -
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. -
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. -
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. -
Sanger Sequencing - Human S100A4 knockout HeLa cell lysate (ab257046)Allele-1: 5 bp deletion in exon2
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Sanger Sequencing - Human S100A4 knockout HeLa cell lysate (ab257046)Allele-2: 1 bp insertion in exon2
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab257046 has not yet been referenced specifically in any publications.