Product nameHuman POLM knockout HeLa cell lysate
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
User storage instructions: After reconstitution, store the lysate at -80°C.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.
Knockout validationSanger Sequencing, Western Blot (WB)
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.
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit Human POLM knockout HeLa cell lysate (Lyophilized) 1 x 100µg Human Wild Type HeLa cell lysate (Lyophilized) 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
FunctionGap-filling polymerase involved in repair of DNA double-strand breaks by non-homologous end joining (NHEJ). Participates in immunoglobulin (Ig) light chain gene rearrangement in V(D)J recombination.
Tissue specificityExpressed in a number of tissues. Abundant in thymus.
Sequence similaritiesBelongs to the DNA polymerase type-X family.
Contains 1 BRCT domain.
- Information by UniProt
- DNA-directed DNA polymerase mu
- DNA-directed DNA/RNA polymerase mu
Our Abpromise guarantee covers the use of ab258127 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 55 kDa.|
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: POLM knockout HeLa cell lysate (20 µg)
Lane 3: HEK-293 cell lysate (20 µg)
Lane 4: Daudi cell lysate (20 µg)
ab157465 Anti-DNA polymerase mu antibody [EPR10470(B)] was shown to specifically react with POLM in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265557 (knockout cell lysate ab258127) was used. Wild-type and POLM knockout samples were subjected to SDS-PAGE. ab157465 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.
Homozygous: 1 bp insertion in exon4
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.
ab258127 has not yet been referenced specifically in any publications.