Human SPOP knockout HeLa cell lysate (ab257702)
Overview
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Product name
Human SPOP knockout HeLa cell lysate
See all SPOP kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HeLa -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon4 and 16 bp deletion in exon4. -
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 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.
Tested applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262194 - Human SPOP knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 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
Inhibits IPF1/PDX1 transactivation of established target promoters, such as insulin, may be by recruiting a repressor complex (By similarity). In complex with CUL3, involved in ubiquitination of BMI1, H2AFY and DAXX, and probably also in ubiquitination and proteasomal degradation of Gli2 or Gli3. -
Tissue specificity
Widely expressed. -
Pathway
Protein modification; protein ubiquitination. -
Sequence similarities
Belongs to the Tdpoz family.
Contains 1 BTB (POZ) domain.
Contains 1 MATH domain. -
Domain
The MATH domain interacts with H2AFY and BMI1. -
Cellular localization
Nucleus. - Information by UniProt
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Alternative names
- BTBD32
- HIB homolog 1
- Roadkill homolog 1
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab262194 - Human SPOP 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
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Western blot - Human SPOP knockout HeLa cell lysate (ab257702)
Lane 1: Wild-type HeLa cell lysate (20 ug)
Lane 2: SPOP knockout HeLa cell lysate (20 ug)
Lane 3: PC-3 cell lysate (20 ug)
Lanes 1-3: Merged signal (red and green). Green - ab192233 observed at 49 kDa. Red - loading control ab7291 observed at 50 kDa.
ab192233 Anti-SPOP antibody [EPR15771] - C-terminal was shown to specifically react with SPOP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265634 (knockout cell lysate ab257702) was used. Wild-type and SPOP knockout samples were subjected to SDS-PAGE. ab192233 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C 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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Sanger Sequencing - Human SPOP knockout HeLa cell lysate (ab257702)Allele-1: 16 bp deletion in exon4
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Sanger Sequencing - Human SPOP knockout HeLa cell lysate (ab257702)Allele-2: 10 bp deletion in exon4