SF20/MYDGF Overexpression Lysate Summary
| Description |
SF20/MYDGF Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC204011
Accession#: NM_019107
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 |
MYDGF |
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 |
18.6 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 SF20/MYDGF Overexpression Lysate
Background
Myeloid-Derived Growth Factor, or MYDGF, is a Bone marrow-derived monocyte protein, and it is correlated with enhanced metabolic activity, suppression of apoptosis, and stimulation of cell proliferation (1). MYDGF is expressed predominantly in inflammatory cells, such as monocytes and macrophages (1). Up-regulation of MYDGF expression was also found during adipocyte differentiation (2). Expression of MYDGF was induced in the circulation and heart tissue after myocardial infarction. It promotes cardiac myocyte survival by stimulating endothelial cell proliferation through a MAPK1/3-, STAT3- and CCND1-mediated signaling pathway, and inhibits cardiac myocyte apoptosis in a PI3K/AKT-dependent signaling pathway (1). MYDGF was found over-expressed in approximately two-thirds of Hepatocellular Carcinoma (HCC) tissues, and its expression was significantly positively correlated with that of alpha-fetoprotein (AFP) (3). In HCC, MYDGF could regulate cell proliferation through activating Akt/mitogen-activated protein kinase pathways (3). Mouse MYDGF shares 92% amino acid sequence identity with both human and rat MYDGF. Intriguingly, virtually all homologs of MYDGF have a C-terminal putative ER retention sequence BXEL (B: Arg, His, or Lys; X: variable residue; E: Glu; L: Leu), which has the potential to retain human MYDGF and its homologs in the ER, whereas truncated MYDGF without BXEL is secreted from the cell (4). However, the functions of these different forms remain unclear.
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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