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 |
|
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
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
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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