Human FHL2 knockout HeLa cell lysate (ab257441)
Overview
-
Product name
Human FHL2 knockout HeLa cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon3 and 1 bp insertion in exon3. -
Passage number
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.
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.
Tested applications
Suitable for: WBmore detailsProperties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260990 - Human FHL2 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
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
-
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
-
Alternative names
- AAG 11
- AAG11
- Aging associated gene 11
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260990 - Human FHL2 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg -
Research areas
-
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
Images
-
Western blot - Human FHL2 knockout HeLa cell lysate (ab257441)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: FHL2 knockout HeLa cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab202586 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.
ab202586 Recombinant Anti-FHL2 antibody [EPR17860-23] was shown to specifically react with FHL2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265475 (knockout cell lysate ab257441) was used. Wild-type and FHL2 knockout samples were subjected to SDS-PAGE. ab202586 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Western blot - Human FHL2 knockout HeLa cell lysate (ab257441)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: FHL2 knockout HeLa cell lysate (20 µg)
Lane 3: K562 cell lysate (20 µg)
Lanes 1-3: Merged signal (red and green). Green - ab202584 observed at 32 kDa. Red - loading control ab7291 observed at 50 kDa.
ab202584 Recombinant Anti-FHL2 antibody [EPR17860-20] was shown to specifically react with FHL2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265475 (knockout cell lysate ab257441) was used. Wild-type and FHL2 knockout samples were subjected to SDS-PAGE. ab202584 and Anti-alpha Tubulin antibody [DM1A] - Loading Control?(ab7291) were incubated overnight at 4^°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
-
Sanger Sequencing - Human FHL2 knockout HeLa cell lysate (ab257441)Allele-1: 1 bp insertion in exon3
-
Sanger Sequencing - Human FHL2 knockout HeLa cell lysate (ab257441)Allele-2: 1 bp deletion in exon3