GNAQ Overexpression Lysate Summary
Description |
GNAQ Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC209199
Accession#: NM_002072
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 |
GNAQ |
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 |
42 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 GNAQ Overexpression Lysate
Background
GNAQ, also known as Guanine nucleotide-binding protein G(q) subunit alpha, is a 359 amino acid that is 42 kDa, with subcellular nucleus location, expressed in ovary, prostate, testis and colon tissue; necessary modulator or transducer in various transmembrane signaling systems; controls B-cell selection and survival, needed to prevent B-cell-dependent autoimmunity; and also regulates chemotaxis of BM-derived neutrophils and dendritic cells. Studies on this protein have shown a relationship with the following diseases and disorders: bleeding diathesis due to gnaq deficiency, cellular blue nevus, pertussis, melanoma, acanthocytosis, fainting, polycystic ovary syndrome, trypanosomiasis, gitelman syndrome chorea, thyroid nodule, huntington's disease, thyroiditis, chagas disease, alzheimer's disease, melanoma metastasis, vasculitis, neuronitis, erythroleukemia, and heart failure. GNAQ has also shown an interaction with AGTR1, CXCR5, CRHR1, GALR2, HTR2C, and over 100 other interacting proteins in many different pathways including calcium signaling pathway, vascular smooth muscle contraction, gap junction, long-term potentiation, glutamatergic synapse, GPCR downstream signaling, hemostasis, gastrin-CREB signalling pathway via PKC and MAPK, signaling by GPCR, signal amplification.
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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