Human LAMTOR3 (MAP2K1IP1) knockout HeLa cell lysate (ab258021)
Overview
-
Product name
Human LAMTOR3 (MAP2K1IP1) 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 insertion in exon5 and 58 bp deletion in exon5. -
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 ab262284 - Human LAMTOR3 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
As part of the Ragulator complex it is involved in amino acid sensing and activation of mTORC1, a signaling complex promoting cell growth in response to growth factors, energy levels, and amino acids. Activated by amino acids through a mechanism involving the lysosomal V-ATPase, the Ragulator functions as a guanine nucleotide exchange factor activating the small GTPases Rag. Activated Ragulator and Rag GTPases function as a scaffold recruiting mTORC1 to lysosomes where it is in turn activated. Adapter protein that enhances the efficiency of the MAP kinase cascade facilitating the activation of MAPK2. -
Sequence similarities
Belongs to the LAMTOR3 family. -
Cellular localization
Late endosome membrane. - Information by UniProt
-
Alternative names
- LAMTOR3
- Late endosomal/lysosomal adaptor and MAPK and MTOR activator 3
- LTOR3_HUMAN
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262284 - Human LAMTOR3 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 LAMTOR3 knockout HeLa cell lysate (ab258021)
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: LAMTOR3 knockout HeLa cell lysate (20 µg)
Lanes 1-2: Merged signal (red and green). Green - ab32134 observed at 14 kDa. Red - loading control ab8245 observed at 37 kDa.
ab32134 Anti-MAP2K1IP1 antibody [Y130] was shown to specifically react with MAP2K1IP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265624 (knockout cell lysate ab258021) was used. Wild-type and MAP2K1IP1 knockout samples were subjected to SDS-PAGE. ab32134 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 50000 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 LAMTOR3 knockout HeLa cell lysate (ab258021)Allele-1: 58 bp deletion in exon5
-
Sanger Sequencing - Human LAMTOR3 knockout HeLa cell lysate (ab258021)Allele-2: 1 bp insertion in exon5