BMP-9 Overexpression Lysate Summary
Description |
BMP-9 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC210204
Accession#: NM_016204
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 |
GDF2 |
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 |
47.1 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 BMP-9 Overexpression Lysate
Background
Bone Morphogenetic Proteins (BMPs) are members of the TGF beta superfamily of cytokines that affect bone and cartilage formation. Similar to other TGF beta family proteins, BMPs are highly conserved across animal species. Mature BMPs are 30-38 kDa proteins that assume a TGF beta like cysteine knot configuration. Unlike TGF beta, BMPs do not form latent complexes with their propeptide counterparts. Most BMPs are homodimers, but bioactive natural heterodimers have been reported. Recently it was found that lovostatin, widely used for lowering cholesterol, also increases bone formation by turning on a gene (bmp2) that promotes local bone formation. BMPs create an environment conducive for bone marrow development by stimulating the production of specific bone matrix proteins and altering stromal cell and osteoclast proliferation. In addition to stimulating ectopic bone and cartilage development, BMPs may be an important factor in the development of the viscera, with roles in cell proliferation, apoptosis, differentiation, and morphogenesis. BMPs also appear to be responsible for normal dorsal/ventral patterning. BMPs are found in tissues that induce bone or cartilage growth, such as demineralized bone and urinary epithelium. Cellular responses to BMP5 are mediated by the formation of hetero-oligomeric complexes of type I and type II serine/threonine kinase receptors which play significant roles in BMP binding and signaling. One BMP type II receptor and two BMP type I receptors have been identified.
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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