Human FHL2 knockout U-2 OS cell lysate (ab263928)
Overview
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Product name
Human FHL2 knockout U-2 OS cell lysate
See all FHL2 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
U-2 OS -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 96.21% -
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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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 ab280481 - Human FHL2 knockout U2OS cell lysate 1 x 100µg ab263974 - Human wild-type U-2 OS cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Osteosarcoma -
Gender
Female
Target
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Function
May function as a molecular transmitter linking various signaling pathways to transcriptional regulation. Negatively regulates the transcriptional repressor E4F1 and may function in cell growth. -
Tissue specificity
Expressed in skeletal muscle and heart. -
Sequence similarities
Contains 4 LIM zinc-binding domains. -
Domain
The third LIM zinc-binding mediates interaction with E4F1. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Alternative names
- AAG 11
- AAG11
- Aging associated gene 11
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 ab263928 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 |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 32 kDa. |
Images
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Next Generation Sequencing - Human FHL2 knockout U-2 OS cell lysate (ab263928)Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift = 96.21%
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Western blot - Human FHL2 knockout U-2 OS cell lysateLane 1: Wild-type U-2 OS cell lysate
Lane 2: FHL2 knockout U-2 OS cell lysate
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysateLanes 1 - 3: Merged signal (red and green). Green - ab202586 observed at 32 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab202586 was shown to react with FHL2 in wild-type U-2 OS cells in western blot with loss of signal observed in FHL2 knockout sample. Wild-type and FHL2 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab202586 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 hour at room temperature before imaging.
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Western blot - Human FHL2 knockout U-2 OS cell lysate (ab263928)Lane 1: Wild-type U-2 OS cell lysate
Lane 2: FHL2 knockout U-2 OS cell lysate
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lanes 1 - 3: Merged signal (red and green). Green - ab202584 observed at 32 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab202584 was shown to react with FHL2 in wild-type U-2 OS cells in western blot with loss of signal observed in FHL2 knockout cell line ab262496 (knockout cell lysate ab263928).Wild-type and FHL2 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab202584 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 hour at room temperature before imaging.
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)
ab263928 has not yet been referenced specifically in any publications.