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cGK1/PRKG1 Overexpression Lysate

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Western Blot: cGK1/PRKG1 Overexpression Lysate (Adult Normal) [NBL1-14782] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for cGK1/PRKG1.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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cGK1/PRKG1 Overexpression Lysate Summary

Description

cGK1/PRKG1 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC220986

Accession#: NM_006258

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
Gene
PRKG1

Applications/Dilutions

Dilutions
  • Western Blot
Application Notes
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW
77.6 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

Storage
Store at -80C. Avoid freeze-thaw cycles.
Buffer
RIPA buffer

Lysate Details for Array

Type
Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for cGK1/PRKG1 Overexpression Lysate

  • AAT8
  • cGK 1
  • cGK
  • cGK1
  • cGK1/PRKG1
  • cGKI
  • cGKI-alpha
  • cGKI-BETA
  • cGMP-dependent protein kinase I
  • DKFZp686K042
  • EC 2.7.11
  • EC 2.7.11.12
  • FLJ36117
  • PKG
  • PKG1
  • PRKG1
  • PRKG1B
  • PRKG1BMGC71944
  • PRKGR1A
  • PRKGR1B
  • PRKGR1BcGMP-dependent, regulatory, type I, beta
  • protein kinase cGMP-dependent 1
  • protein kinase, cGMP-dependent, type I

Background

cGK1/PRKG1, also known as cGMP-dependent protein kinase 1, has two isoforms that are produced by alternative splicing. Isoform Alpha (CGK1-alpha) is 671 amino acids long and 76 kDa. Isoform Beta (CGK1-beta) is 686 amino acids long and approximately 78 kDa. PRKG1 is a Serine/threonine protein kinase that is expressed in placenta and lung. cGK1/PRKG1 is an important player in the Nitric Oxide/cGMP Signaling Pathway; the binding of GMP activates PRKG1, which in turn phosphorylates Serine residues and Threonine residues on various cell proteins, including proteins that regulate calcium levels, smooth muscle contraction, platelet adhesion, circadian rhythm and more. Current research on PRKG1 is being conducted in relation to several diseases and disorders including cystic fibrosis, lymphoma, non-small cell lung carcinoma, Wiskott-Aldrich syndrome, Waterhouse-Friderichsen syndrome and myoglobinuria. PRKG1 has also been shown to have interactions with SMAD4, BMPR2, GKAP1, Natriuretic Peptide Receptor A and TFII-I in pathways such as Gap Junction signaling, eNOS signaling and EDNRB signaling.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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Bioinformatics

Gene Symbol PRKG1