Angiopoietin-like Protein 3/ANGPTL3 Overexpression Lysate Summary
Description |
Angiopoietin-like Protein 3/ANGPTL3 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC208867
Accession#: NM_014495
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 |
ANGPTL3 |
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 |
51.4 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 Angiopoietin-like Protein 3/ANGPTL3 Overexpression Lysate
Background
ANGPTL3 is a 460 amino acid protein that is found most commonly in the liver, but has also been shown to reside in kidney samples, and contains three domains consisting of a signal peptide, N-terminal coiled-coil and C-terminal Fibrinogen (FBN) like domain. This protein is involved with the lipid metabolism disease pathways in relation to low levels of LDL (low density lipoproteins), HDL (High density lipoproteins), and triglycerides in plasma. This protein plays a role in angiogenesis and has been shown to interact with ITGAV, ITGB3, and ITGA5, and work is being done to study the protein and the related hypobetalipoproteinemia and atherosclerosis involvement.
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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