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AB257608

Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate

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PRKCSH KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. 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

6 Images
Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)
  • WB

Lab

Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : PRKCSH knockout HEK-293T cell lysate (20µg)

Lane 3 : HeLa cell lysate (20µg)

Lane 4 : Daudi cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab134071 observed at 80 kDa. Red - loading control ab8245 observed at 37 kDa.

ab134071 Anti-Glucosidase 2 subunit beta antibody [EPR8046] was shown to specifically 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 and PRKCSH knockout samples 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) 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.

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 (<a href='/en-us/products/cell-lines/human-prkcsh-glucosidase-2-subunit-beta-knockout-hek-293t-cell-line-ab266770'>ab266770</a>)

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

Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)
  • WB

Lab

Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : PRKCSH knockout HEK-293T cell lysate (20µg)

Lane 3 : HeLa cell lysate (20µg)

Lane 4 : Daudi cell lysate (20µg)

Lanes 1- 4 : Merged signal (red and green). Green - ab129098 observed at 80 kDa. Red - loading control ab8245 observed at 37 kDa.

ab129098 Anti-Glucosidase 2 subunit beta antibody [EPR8047] was shown to specifically 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 and PRKCSH knockout samples 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) were incubated overnight at 4°C at 1 in 10000 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.

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 (<a href='/en-us/products/cell-lines/human-prkcsh-glucosidase-2-subunit-beta-knockout-hek-293t-cell-line-ab266770'>ab266770</a>)

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 Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)
  • Sanger seq

Unknown

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)

Allele-2 : 13 bp deletion in exon 1

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)
  • Sanger seq

Unknown

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)

Allele-3 : Insertion of the selection cassette in exon 1

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)
  • Sanger seq

Unknown

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)

Allele-4 : Insertion of the selection cassette in exon 1

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)
  • Sanger seq

Unknown

Sanger Sequencing - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell lysate (AB257608)

Allele-1 : 14 bp deletion in exon 1

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

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.

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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. 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.

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.

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What's included?

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Properties and storage information

Gene name
PRKCSH
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Glucosidase 2 subunit beta also known as PRKCSH plays an important role in protein processing within the endoplasmic reticulum (ER). This protein forms part of the heterodimeric enzyme complex glucosidase II responsible for cleaving glucose molecules during glycoprotein maturation. With a molecular weight of about 60 kDa glucosidase 2 subunit beta is important for efficient N-linked glycosylation which is a fundamental process for protein folding and quality control. This protein localizes mainly to the ER where it contributes to proper cellular functions by aiding in the preparation of proteins for trafficking to their final destinations.
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.

Defects in glucosidase 2 subunit beta have links to autosomal dominant polycystic liver disease (PCLD). Mutations in PRKCSH which encodes glucosidase 2 subunit beta can lead to the development of PCLD due to disrupted glycoprotein folding and ER stress-induced apoptosis. Additionally abnormalities in its function may indirectly connect to diabetes mellitus by affecting insulin receptor glycosylation and secretion processes. These relationships illustrate the broader impact of glucosidase 2 subunit beta on health and disease providing insight into potential therapeutic targets for treatment strategies.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Complementary Products

Primary Antibodies

AB248669

Anti-Glucosidase 2 subunit beta antibody [EPR8046] - BSA and Azide free

RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Glucosidase 2 subunit beta antibody [EPR8046] - BSA and Azide free (AB248669)

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Cell Lines & Lysates

AB266770

Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line

Advanced Validation

Western blot - Human PRKCSH (Glucosidase 2 subunit beta) knockout HEK-293T cell line (AB266770)

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Product promise

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