Hyaluronan synthase 1 Overexpression Lysate Summary
Description |
Hyaluronan synthase 1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC207797
Accession#: NM_001523
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 |
HAS1 |
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 |
64.7 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 Hyaluronan synthase 1 Overexpression Lysate
Background
Hyaluronan or hyaluronic acid (HA) is a high molecular weight unbranched polysaccharide synthesized by a wide varietyof organisms from bacteria to mammals, and is a constituent of the extracellular matrix. It consists of alternatingglucuronic acid and N-acetylglucosamine residues that are linked by beta-1-3 and beta-1-4 glycosidic bonds. HA issynthesized by membrane-bound synthase at the inner surface of the plasma membrane, and the chains are extrudedthrough pore-like structures into the extracellular space. It serves a variety of functions, including space filling,lubrication of joints, and provision of a matrix through which cells can migrate. HA is actively produced during woundhealing and tissue repair to provide a framework for ingrowth of blood vessels and fibroblasts. Changes in the serumconcentration of HA are associated with inflammatory and degenerative arthropathies such as rheumatoid arthritis. Inaddition, the interaction of HA with the leukocyte receptor CD44 is important in tissue-specific homing by leukocytes,and overexpression of HA receptors has been correlated with tumor metastasis. HAS1 is a member of the newly identifiedvertebrate gene family encoding putative hyaluronan synthases, and its amino acid sequence shows significant homologyto the hasA gene product of Streptococcus pyogenes, a glycosaminoglycan synthetase (DG42) from Xenopus laevis, and arecently described murine hyaluronan synthase. (provided by RefSeq)
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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