Serpin A7/TBG Overexpression Lysate Summary
Description |
Serpin A7/TBG Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC204733
Accession#: NM_000354
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 |
SERPINA7 |
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 |
46.1 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 Serpin A7/TBG Overexpression Lysate
Background
Thyroxine Binding Globulin (TBG) is one of three proteins responsible for carrying the thyroid hormones thyroxine (T4) and 3,5,3 triiodothyronine (T3) in the bloodstream, along with transthyretin and albumin. TBG has the highest affinity for T4 and T3, but is present in the lowest concentration of the three proteins. Despite its low concentration, TBG carries the majority of T4 in serum. Due to the very low serum concentration of T4 & T3, TBG is rarely more than 25% saturated with ligand. Unlike transthyretin and albumin, TBG has a single binding site for T4/T3. Genomically, TBG is a serpin, although it has no inhibitory function like many other members of this class of proteins. TBG is synthesized primarily in the liver.
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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