TFIP11 Overexpression Lysate Summary
Description |
TFIP11 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC200290
Accession#: NM_001008697
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 |
TFIP11 |
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 |
96.6 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 TFIP11 Overexpression Lysate
Background
Neurotensin (NT) initiates an intracellular response by interacting with the G protein-coupled receptors NTR1 (NTS1 receptor, high affinity NTR) and NTR2 (NTS2 receptor, levocabastine-sensitive neurotensin receptor), and the type I receptor NTR3 (NTS3 receptor, sortilin-1, Gp95). NT has a wide distribution in regions of the brain and in peripheral tissues where NT receptors can contribute to hypotension, hyperglycemia, hypothermia, antinociception and regulation of intestinal motility and secretion. HL-60 cells express NTR1, which can couple to Gq, Gi/o, or Gs. Alternative splicing of rat NTR2 can generate a 5-transmembrane domain variant isoform that is co-expressed with the full-length NTR2 throughout the brain and spinal cord. NTR3 activation in the murine microglial cell line N11 induces MIP-2, MCP-1, IL-1beta and TNFalpha in an ERK1/2 and Akt kinase-dependent manner.
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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