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GCAP3 Overexpression Lysate

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Western Blot: GCAP3 Overexpression Lysate (Adult Normal) [NBP2-10466] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for GCAP3.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

Order Details

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GCAP3 Overexpression Lysate Summary

Description

GCAP3 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC216167

Accession#: NM_005459

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
GUCA1C

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
23.6 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 GCAP3 Overexpression Lysate

  • GCAP3GCAP 3
  • guanylate cyclase activator 1CMGC120159
  • guanylyl cyclase-activating protein 3
  • MGC120158

Background

GCAP3, also known as GUCA1C or Guanylyl cyclase-activating protein 3, has a 209 amino acid isoform that is 24 kDa and a short 196 amino acid isoform that is approx. 22 kDa; commonly found in retina; in conditions of low free calcium ions concentration stimulates guanylyl cyclase 1 (GC1) and GC2 and when the concentration is elevated inhibits guanylyl cyclases, which is very important for the recuperation of the dark state of rod photoreceptors following light exposure. Disease research has linked this protein with cone dystrophy, neuronitis, and retinitis. This protein has shown interactions with GUCA1A, GUCA1B, GUCY2D, GUCY2F, and CNGA3 proteins in the pathways such as the visual cycle in retinal rods, olfactory transduction, and phototransduction pathways.

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 GUCA1C