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
LRP6
Applications/Dilutions
Dilutions
Western Blot
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
178.4 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.
Reviewed Applications
Read 1 Review rated 5 using NBL1-12672 in the following application:
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 LRP-6 Overexpression Lysate
ADCAD2
FLJ90062
FLJ90421
low density lipoprotein receptor-related protein 6
low-density lipoprotein receptor-related protein 6
LRP6
LRP-6
Background
Please note, this product is one of a range of Investigative Grade antibodies, made against targets that have limited or no commercial antibodies available to them and for which there are no data on the expression of the protein in the range of common cell lines and tissues available to us. These antibodies are affinity purified using their peptide immunogen and are known to give low background staining in a western blot (see Application Notes below). However no additional claims are made for their ability to recognise native protein in any application. LRP6 is essential for the Wnt/beta catenin signaling pathway, probably by acting as a coreceptor together with Frizzled for Wnt. It is a specific, high-affinity receptor for DKK1 and DKK2, but not DKK3. The interaction with DKK1 blocks LRP6-mediated Wnt/beta catenin signaling via LRP6 removal via Kremen proteins-mediated endocytosis.
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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