Human SCAMP1 knockout HeLa cell lysate (ab258184)
Overview
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Product name
Human SCAMP1 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, 1 bp insertion in exon2 and 25 bp deletion in exon2. -
Passage number
<20 -
Knockout validation
Sanger Sequencing -
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: 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.
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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 ab262336 - Human SCAMP1 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
Functions in post-Golgi recycling pathways. Acts as a recycling carrier to the cell surface. -
Tissue specificity
Widely expressed, with highest expression in brain. -
Sequence similarities
Belongs to the SCAMP family. -
Cellular localization
Golgi apparatus > trans-Golgi network membrane. Recycling endosome membrane. - Information by UniProt
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Alternative names
- SCAM1_HUMAN
- SCAMP
- SCAMP 1
see all
Associated products
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KO cell lines
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab258184 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 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 38 kDa.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 38 kDa. |
Images
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Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: SCAMP1 knockout HeLa cell lysate 20 μgLanes 1 - 2: Merged signal (red and green). Green - Anti-SCAMP1 antibody observed at 36 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa. Anti-SCAMP1 antibody was shown to react with SCAMP1 in wild-type HeLa cells in western blot. The bands observed in SCAMP1 knockout cell line ab265567 (SCAMP1 knockout cell lysate ab258184) below 36 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. HeLa wild-type and SCAMP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with anti-SCAMP1 antibody and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Allele-1: 25 bp deletion in exon2
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Allele-2: 1 bp insertion 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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SDS download
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Datasheet download
References (0)
ab258184 has not yet been referenced specifically in any publications.