Human ULK3 knockout HEK-293T cell lysate (ab258271)
Overview
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Product name
Human ULK3 knockout HEK-293T cell lysate -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9. -
Parental Cell Line
HEK293T -
Organism
Human -
Mutation description
Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 1 bp insertion in exon1. -
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, 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 applications
Suitable for: WBmore detailsProperties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab261194 - Human ULK3 knockout HEK293T cell lysate 1 x 100µg ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg -
Research areas
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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
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Function
Serine/threonine protein kinase which enhances GLI1 and GLI2 transcriptional activity and consequently positively regulates GLI-dependent SHH signaling. May exert this function by promoting GLI1 nuclear localization. Phosphorylates in vitro GLI2, as well as GLI1 and GLI3, although less efficiently. -
Tissue specificity
Widely expressed. Highest levels observed in fetal brain. In adult tissues, high levels in brain, liver and kidney, moderate levels in testis and adrenal gland and low levels in heart, lung, stomach, thymus, prostate and placenta. In the brain, highest expression in the hippocampus, high levels also detected in the cerebellum, olfactory bulb and optice nerve. In the central nervous system, lowest levels in the spinal cord. -
Sequence similarities
Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. APG1/unc-51/ULK1 subfamily.
Contains 2 MIT domains.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated in vitro. -
Cellular localization
Cytoplasm. - Information by UniProt
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Form
There are 3 isoforms produced by alternative splicing. -
Alternative names
- DKFZP434C131
- FLJ90566
- Serine/threonine-protein kinase ULK3
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab261194 - Human ULK3 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
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Western blot - Human ULK3 knockout HEK293T cell lysate (ab258271)
Lane 1:Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 2:ULK3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate (20 ug)
Lane 3:HAP1 whole cell lyate (20 ug)
Lane 4:LNCaP (Human prostate cancer cell line) whole cell lysate (20 ug)ab124947 was shown to specifically react with ULK3 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266152 (knockout cell lysate ab258271) was used. Wild-type and ULK3 knockout samples were subjected to SDS-PAGE. ab124947 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.
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Sanger Sequencing - Human ULK3 knockout HEK293T cell lysate (ab258271)Allele-1: 1 bp deletion in exon1
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Sanger Sequencing - Human ULK3 knockout HEK293T cell lysate (ab258271)Allele-2: 1 bp insertion in exon1