Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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PRKCSH KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 13 bp deletion in exon 1 and 14 bp deletion in exon 1 and Insertion of the selection cassette in exon 1.
View Alternative Names
80K-H protein, AGE-R2, AGE-binding receptor 2, G19P1, GLU2B_HUMAN, Glucosidase 2 subunit beta, Glucosidase II beta subunit, Glucosidase II subunit beta, Hepatocystin, PCLD, PKCSH, PRKCSH, Protein kinase C substrate 60.1 kDa protein heavy chain, Protein kinase C substrate 80 Kda protein, Protein kinase C substrate 80K-H
- WB
Lab
Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)
Lanes 1- 4 : Merged signal (red and green). Green - ab129098 observed at 80 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab129098 was shown to react with PRKCSH in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266770 (knockout cell lysate ab257608) was used. Wild-type HEK-293T and PRKCSH knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab129098 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 10000 dilution and a 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.
All lanes:
Western blot - Anti-Glucosidase 2 subunit beta antibody [EPR8047] (<a href='/en-us/products/primary-antibodies/glucosidase-2-subunit-beta-antibody-epr8047-ab129098'>ab129098</a>) at 1/10000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PRKCSH knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (ab266770)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 59 kDa
Observed band size: 80 kDa
false
- WB
Lab
Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)
Lanes 1- 4 : Merged signal (red and green). Green - ab134071 observed at 80 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab134071 was shown to react with PRKCSH in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266770 (knockout cell lysate ab257608) was used. Wild-type HEK-293T and PRKCSH knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab134071 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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.
All lanes:
Western blot - Anti-Glucosidase 2 subunit beta antibody [EPR8046] (<a href='/en-us/products/primary-antibodies/glucosidase-2-subunit-beta-antibody-epr8046-ab134071'>ab134071</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
PRKCSH knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (ab266770)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 59 kDa
Observed band size: 80 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)
Allele-2 : 13 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)
Allele-3 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)
Allele-4 : Insertion of the selection cassette in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)
Allele-1 : 14 bp deletion in exon 1
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Glucosidase 2 subunit beta is associated with protein maturation and quality control within the ER lumen. It is part of the glucosidase II complex which functions to trim glucose residues from N-linked oligosaccharides on nascent glycoproteins facilitating their proper folding and preventing misfolded proteins' accumulation. This activity ensures that only properly folded proteins progress through the secretory pathway while faulty proteins are targeted for degradation. Through these processes glucosidase 2 subunit beta supports cellular homeostasis and protein turnover.
Pathways
Glucosidase 2 subunit beta plays an influential role in the protein processing and ER-associated degradation (ERAD) pathways. The protein operates within these pathways to regulate glycoprotein folding impacting the unfolding protein response (UPR). It interacts with several proteins including calnexin and calreticulin which are key players in the glycoprotein folding process. Moreover the proper function of glucosidase 2 subunit beta in these pathways is necessary for maintaining the cellular stress response ultimately ensuring efficient cell function and survival.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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