Human GOLIM4 (GOLPH4/GPP130) knockout HEK-293T cell lysate (ab257455)
Overview
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Product name
Human GOLIM4 (GOLPH4/GPP130) 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, Homozygous: 2 bp insertion in exon 1. -
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 ab260992 - Human GOLIM4 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
Plays a role in endosome to Golgi protein trafficking; mediates protein transport along the late endosome-bypass pathway from the early endosome to the Golgi. -
Sequence similarities
Belongs to the GOLIM4 family. -
Post-translational
modificationsPhosphorylated probably by c-AMP-dependent kinases in its lumenal part.
O-glycosylated; modified by sialic acid residues.
N-glycosylated; N-glycans are probably of the complex type and modified by sialic acid residues. -
Cellular localization
Golgi apparatus > Golgi stack membrane. Endosome membrane. Localizes to cis and medial Golgi cisternae. Probably cycles between early Golgi and distal compartments like endosome. - Information by UniProt
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Alternative names
- 130 kDa golgi localized phosphoprotein
- cis
- cis Golgi localized calcium binding protein
see all
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab260992 - Human GOLIM4 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 GOLIM4 knockout HEK293T cell lysate (ab257455)
Lane 1: Wild-type HEK293T cell lysate (20 µg)
Lane 2: GOLIM4 knockout HEK293T cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: MOLT-4 cell lysate (20 µg)
Lanes 1-4: Merged signal (red and green). Green - ab197595 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.
ab197595 Anti-GOLPH4/GPP130 antibody [EPR13439] was shown to specifically react with GOLPH4/GPP130 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266123 (knockout cell lysate ab257455) was used. Wild-type and GOLPH4/GPP130 knockout samples were subjected to SDS-PAGE. ab197595 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.
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Sanger Sequencing - Human GOLIM4 knockout HEK293T cell lysate (ab257455)Homozygous: 2 bp insertion in exon 1
