Human VIM (Vimentin) knockout HeLa cell pellet (ab278921)
Overview
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Product name
Human VIM (Vimentin) knockout HeLa cell pellet
See all Vimentin kits -
Product overview
Abcam’s knockout cell pellets give you access to native proteins, without the need to culture cells. Our knockout cell pellets are prepared from our single-gene knockout cell lines and provide an additional offering to our cell lysates.
Cells are snap-frozen to provide high quality pellets that are suitable for extraction with alternative lysis buffers or for preparation of lysates from subcellular fractions. Our knockout cell pellets are suitable for a variety of applications, including PCR, gene expression profiling and DNA library preparation. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing, Western Blot (WB) -
Notes
Pellet size: 5 million cells/vial.
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.
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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 Human VIM knockout HeLa cell pellet 1 vial Human wild-type HeLa cell pellet 1 vial -
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
Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2. -
Tissue specificity
Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines. -
Involvement in disease
Cataract 30 -
Sequence similarities
Belongs to the intermediate filament family. -
Domain
The central alpha-helical coiled-coil rod region mediates elementary homodimerization.
The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex. -
Post-translational
modificationsFilament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33.
O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.
S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex. -
Cellular localization
Cytoplasm. - Information by UniProt
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Form
Vimentin is found in connective tissue and in the cytoskeleton. -
Alternative names
- CTRCT30
- Epididymis luminal protein 113
- FLJ36605
see all
Associated products
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KO cell lines
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KO cell lysates
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab278921 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: 53 kDa.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa. |
Images
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Western blot - Human VIM (Vimentin) knockout HeLa cell pellet (ab278921)
Lane 1: U20S cell lysate (20 µg)
Lane 2: Hu tonsil cell lysate (20 µg)
Lane 3: Wild-type HeLa cell lysate (20 µg)
Lane 4: VIM knockout HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab8069 observed at 53 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab8069 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab8069 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 µg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Human VIM (Vimentin) knockout HeLa cell pellet (ab278921)Lane 1: U20S cell lysate (20 µg)
Lane 2: Hu tonsil cell lysate (20 µg)
Lane 3: Wild-type HeLa cell lysate (20 µg)
Lane 4: VIM knockout HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab16700 observed at 53 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab16700 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab16700 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 100 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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Sanger Sequencing - Human VIM (Vimentin) knockout HeLa cell pellet (ab278921)Homozygous: Insertion of the selection cassette in exon2
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.
Datasheets and documents
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Datasheet download
References (0)
ab278921 has not yet been referenced specifically in any publications.
