Human NMI (N myc interactor/NMI) knockout A549 cell lysate (ab258077)
Overview
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Product name
Human NMI (N myc interactor/NMI) knockout A549 cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon5 and 330 bp insertion 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
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262297 - Human NMI 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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Relevance
NMYC interactor (NMI) encodes a protein that interacts with NMYC and CMYC (two members of the oncogene Myc family), and other transcription factors containing a Zip, HLH, or HLH Zip motif. The NMI protein also interacts with all STATs except STAT2 and augments STAT mediated transcription in response to cytokines IL2 and IFN gamma. The NMI mRNA has high expression in myeloid leukemia cell lines. -
Cellular localization
Cytoplasmic -
Alternative names
- N myc (and STAT) interactor
- N myc and STAT interactor
- N myc interactor
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262297 - Human NMI 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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Lane 1:Wild-type A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)
Lane 2:NMI knockout A549 (Human lung carcinoma cell line) whole cell lysate (20 ug)
Lane 3:HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate (20 ug)
Lane 4:K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate (20 ug)ab183724 was shown to specifically react with N myc interactor/NMI in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267013 (knockout cell lysate ab258077) was used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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.
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Allele-1: 2 bp deletion in exon5
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Allele-2: 330 bp insertion in exon5