Human S100A4 knockout A549 cell lysate (ab261674)
Overview
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Product name
Human S100A4 knockout A549 cell lysate
See all S100A4 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 95% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), 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: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280424 - Human S100A4 knockout A549 cell lysate 1 x 100µg ab259782 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
Gender
Male
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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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab261674 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 |
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WB |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. |
Images
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Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 μg
Lane 2: S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate 20 μg
Lane 3: A-375 (Human malignant melanoma cell line) whole cell lysate 20 μg
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 μg
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. -
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 2: S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 3: A-375 (Human malignant melanoma cell line) whole cell lysate 20 ug
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
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. -
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 2: S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 3: A-375 (Human malignant melanoma cell line) whole cell lysate 20 ug
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
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. -
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 2: S100A4 knockout A549 (Human lung carcinoma cell line) whole cell lysate 20 ug
Lane 3: A-375 (Human malignant melanoma cell line) whole cell lysate 20 ug
Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate 20 ug
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. -
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 95%
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)
ab261674 has not yet been referenced specifically in any publications.