Human DAZAP1 knockout HEK-293T cell lysate (ab257912)
Overview
-
Product name
Human DAZAP1 knockout HEK-293T cell lysate
See all DAZAP1 kits -
Product overview
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
-
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 8 bp deletion in exon 6. -
Passage number
Knockout validation
Sanger SequencingReconstitution 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
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260389 - Human DAZAP1 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Target
-
Function
RNA-binding protein, which may be required during spermatogenesis. -
Tissue specificity
Mainly expressed in testis. Expressed to a lower level in thymus. Weakly or not expressed in heart, liver, brain, placenta, lung, skeletal muscle, kidney and pancreas. -
Sequence similarities
Contains 2 RRM (RNA recognition motif) domains. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Cytoplasm. Nucleus. Predominantly cytoplasmic (By similarity). Nuclear at some stages of spermatozoides development. In midpachytene spermatocytes, it is localized in both the cytoplasm and the nuclei and is clearly excluded from the sex vesicles. In round spermatids, it localizes mainly in the nuclei, whereas in elongated spermatids, it localizes to the cytoplasm. - Information by UniProt
-
Alternative names
- DAZ associated protein 1
- DAZ-associated protein 1
- Dazap1
see all
Properties
-
Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260389 - Human DAZAP1 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
-
Cell type
epithelial -
STR Analysis
Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12
Images
-
Western blot - Human DAZAP1 knockout HEK293T cell lysate (ab257912)
Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:DAZAP1 knockout HEK293T cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)
Lane 4:HeLa cell lysate (20 ug)ab184183 was shown to specifically react with DAZAP1 in wild-type HEK293T cells. The band observed in knockout cell line ab266469 (knockout cell lysate ab257912) lane below 43 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and DAZAP1 knockout samples were subjected to SDS-PAGE. ab184183 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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.
-
Sanger Sequencing - Human DAZAP1 knockout HEK293T cell lysate (ab257912)Homozygous: 8 bp deletion in exon 6
