Surfactant Protein A Overexpression Lysate Summary
Description |
Surfactant Protein A Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC228781
Accession#: NM_001164644
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 |
SFTPA1 |
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 |
26.1 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 48 hrs before collection. The cells were lysed in modified RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1 mM EDTA, 1X Proteinase inhibitor cocktail mix, 1 mM PMSF and 1 mM Na3PO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for Surfactant Protein A Overexpression Lysate
Background
Surfactant protein A is a protein with two isoforms, with lengths of 248 and 263 amino acids and weights of approximately 26 and 28 kDa respectively. Surfactant protein A binds to surfactant phospholipids in the presence of calcium ions and helps lower the surface tension in the alveoli of the lungs. Current research is being done on several diseases and disorders linked to this protein including pulmonary fibrosis, allergic bronchopulmonary aspergillosis, pulmonary alveolar proteinosis, non-small cell lung carcinoma, sclerosing hemangioma, otitis media, chronic obstructive pulmonary disease, bronchitis obliterans, bronchopulmonary dysplasia, rhinitis, aspergillosis, pulmonary edema, insulin resistance, and pneumoconiosis. Surfactant protein A has also been shown to have interactions with DMBT1, MYO18A, C1QA, SFTPD, and TLR2 in pathways such as the immune response, bacterial infections in CF airways, cell-cell communication, phagosome, and pertussis pathways.
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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