Human MVP knockout HeLa cell lysate (ab257544)
Overview
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Product name
Human MVP 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, 17 bp deletion in exon 6 and 1 bp deletion in exon 6. -
Passage number
<20 -
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.
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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: After reconstitution, store the lysate at -80°C.
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 ab260277 - Human MVP knockout HeLa cell lysate (Lyophilized) 1 x 100µg ab255552 - Human wild-type HeLa cell lysate (Lyophilized) 1 x 100µg -
Research areas
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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
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Function
Required for normal vault structure. Vaults are multi-subunit structures that may act as scaffolds for proteins involved in signal transduction. Vaults may also play a role in nucleo-cytoplasmic transport. Down-regulates INFG-mediated STAT1 signaling and subsequent activation of JAK. Down-regulates SRC activity and signaling through MAP kinases. -
Tissue specificity
Present in most normal tissues. Higher expression observed in epithelial cells with secretory and excretory functions, as well as in cells chronically exposed to xenobiotics, such as bronchial cells and cells lining the intestine. Overexpressed in many multidrug-resistant cancer cells. -
Sequence similarities
Contains 9 MVP (vault) repeats. -
Domain
MVP 3 mediates interaction with PTEN.
MVP 4 mediates interaction with PARP4. -
Post-translational
modificationsPhosphorylated on Tyr residues after EGF stimulation.
Dephosphorylated by PTPN11. -
Cellular localization
Cytoplasm. Nucleus > nuclear pore complex. 5% found in the nuclear pore complex. Translocates from the nucleus to the cytoplasm upon EGF treatment. - Information by UniProt
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Alternative names
- LRP
- Lung resistance related protein
- Lung resistance-related protein
see all
Associated products
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KO cell lines
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Related Products
Applications
Our Abpromise guarantee covers the use of ab257544 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: 99 kDa. |
Images
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Western blot - Human MVP knockout HeLa cell lysate (ab257544)
Lane 1: Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate (20 ug)
Lane 2: MVP knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate (20 ug)
Lane 3: Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate (20 ug)
Lanes 1-3: Merged signal (red and green). Green - ab273093 observed at 110 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.
ab273093 Anti-MVP antibody [EPR23594-106] was shown to specifically react with MVP in wild-type HeLa cells in Western blot. Significant decrease (8.7 % of intensity compared to the WT band) of signal was observed when MVP knockout cell line ab264817 (knockout cell lysate ab257544) was used.
ab273093 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4? overnight 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 10000 dilution for 1 hour at room temperature before imaging. -
Western blot - Human MVP knockout HeLa cell lysate (ab257544)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: MVP knockout HeLa cell lysate (20 µg)
Lane 3: A549 cell lysate (20 µg)
Lane 4: MOLT-4 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab177145 observed at 99 kDa. Red - loading control ab8245 observed at 37 kDa.
ab177145 Anti-MVP antibody [EPR13226(B)] was shown to specifically react with MVP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264817 (knockout cell lysate ab257544) was used. Wild-type and MVP knockout samples were subjected to SDS-PAGE. ab177145 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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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Western blot - Human MVP knockout HeLa cell lysate (ab257544)Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: MVP knockout HeLa cell lysate (20 µg)
Lane 3: A549 cell lysate (20 µg)
Lane 4: MOLT-4 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab175239 observed at 110 kDa. Red - loading control ab8245 observed at 37 kDa.
ab175239 Anti-MVP antibody [EPR13227(B)] was shown to specifically react with MVP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264817 (knockout cell lysate ab257544) was used. Wild-type and MVP knockout samples were subjected to SDS-PAGE. ab175239 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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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Sanger Sequencing - Human MVP knockout HeLa cell lysate (ab257544)Allele-1: 17 bp deletion in exon 6
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Sanger Sequencing - Human MVP knockout HeLa cell lysate (ab257544)Allele-2: 1 bp deletion in exon 6
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.
References (0)
ab257544 has not yet been referenced specifically in any publications.
