Human MAP3K2 (MEKK2) knockout A549 cell lysate (ab257522)
Overview
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Product name
Human MAP3K2 (MEKK2) knockout A549 cell lysate
See all MEKK2 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon7 and 4 bp deletion in exon7. -
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
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262113 - Human MAP3K2 knockout A549 cell lysate 1 x 100µg ab255554 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X,Y D5S818: 11 D13S317: 11 D7S820: 8, 11 D16S539: 11, 12 vWA: 14 TH01: 8,9.3 TPOX: 8,11 CSF1PO: 10, 12
Target
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Function
Component of a protein kinase signal transduction cascade. Regulates the JNK and ERK5 pathways by phosphorylating and activating MAP2K5 and MAP2K7 (By similarity). Plays a role in caveolae kiss-and-run dynamics. -
Sequence similarities
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
Contains 1 OPR domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated. -
Cellular localization
Cytoplasm. Nucleus. Upon EGF stimulation, translocates into the nucleus. - Information by UniProt
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Alternative names
- M3K2_HUMAN
- Map3k2
- MAPK/ERK kinase kinase 2
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262113 - Human MAP3K2 knockout A549 cell lysate 1 x 100µg ab255554 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
Disease
Carcinoma -
STR Analysis
Amelogenin X,Y D5S818: 11 D13S317: 11 D7S820: 8, 11 D16S539: 11, 12 vWA: 14 TH01: 8,9.3 TPOX: 8,11 CSF1PO: 10, 12
Images
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Western blot - Human MAP3K2 (MEKK2) knockout A549 cell lysate (ab257522)Lane 1: Wild-type A549 cell lysate (20µg)
Lane 2: MAP3K2 knockout A549 cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab33918 observed at 75 kDa. Red - loading control ab8245 observed at 37 kDa.
ab33918 Anti-MEKK2 antibody [EP626Y] was shown to specifically react with MEKK2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267153 (knockout cell lysate ab257522) was used. Wild-type and MEKK2 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab33918 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 µg/ml 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 MAP3K2 knockout A549 cell lysate (ab257522)Allele-1: 4 bp deletion in exon7
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Sanger Sequencing - Human MAP3K2 knockout A549 cell lysate (ab257522)Allele-2: 1 bp insertion in exon7
