Product nameHuman SERPINB5 (MASPIN) knockout HeLa cell lysate
See all MASPIN kits
Knockout cell lysate achieved by CRISPR/Cas9.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
Knockout validationSanger Sequencing
Reconstitution notesTo 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.
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.
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit ab261308 - Human SERPINB5 knockout HeLa cell lysate 1 x 100µg ab255929 - Human wild-type HeLa cell lysate 1 x 100µg
STR AnalysisAmelogenin 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
FunctionTumor suppressor. It blocks the growth, invasion, and metastatic properties of mammary tumors. As it does not undergo the S (stressed) to R (relaxed) conformational transition characteristic of active serpins, it exhibits no serine protease inhibitory activity.
Tissue specificityNormal mammary epithelial cells.
Sequence similaritiesBelongs to the serpin family. Ov-serpin subfamily.
Cellular localizationSecreted > extracellular space.
- Information by UniProt
- Peptidase inhibitor 5
KO cell lines
Lane 1: Wild-type HeLa cell lysate 20 μg
Lane 2: SERPINB5 knockout HeLa cell lysate 20 μg
Lane 3: HaCaT cell lysate 20 μg
Lane 4: Caco-2 cell lysate 20 μg
False colour image of Western blot: Anti-MASPIN antibody (ab272858) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab272858 was shown to bind specifically to MASPIN. A band was observed at 42 kDa in wild-type HeLa cell lysates with no signal observed at this size in SERPINB5 knockout cell line ab264750 (knockout cell lysate ab258659). To generate this image, wild-type and SERPINB5 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Homozygous: 1 bp insertion in exon 2
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.
ab258659 has not yet been referenced specifically in any publications.