TUBGCP5 Overexpression Lysate Summary
Description |
TUBGCP5 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC209703
Accession#: NM_052903
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 |
TUBGCP5 |
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 |
118.1 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 TUBGCP5 Overexpression Lysate
Background
The gamma-Tubulin complex is composed of gamma Tubulin and the gamma-Tubulin complexassociated proteins GCP2, GCP3, GCP4, GCP5 and GCP6, all of which are essential components of microtubule organizing centers. gamma-Tubulin complex components are localized to both the centrosome, where they are involved in microtubule nucleation, and to the cytoplasm, where they exist as soluble complexes that can be recruited to the centrosome as needed. Although the GCP proteins are related, they have distinct roles which contribute to the proper function of the gamma-Tubulin complex. GCP5 (gamma-Tubulin complex component 5), also known as TUBGCP5, is a 1,024 amino acid member of the gamma-Tubulin complex and is highly expressed in heart and skeletal muscle. Defects in the gene encoding GCP5 are implicated in Prader-Willi syndrome (PWS), a rare genetic disorder associated with obesity, compulsive behavior and lower intellectual ability.
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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