Type
Native
Description
Native protein isolated from pooled human serum 226 AA. MW: 24.54 kDa (calculated without glycosylation). Protein identity confirmed by LC-MS/MS. Identification of Adiponectin from human serum by LC-MS/MS – The 60kDa and 37 kDa bands from SDS-PAGE were excised from the polyacrylamide gel and in-gel digested. Tryptic peptides were analyzed by LC-MS/MS and the both bands were identified as adiponectin precursor (NCBI Reference Sequence: NP_004788.1).
Amino Acid Sequence
ETTTQGPGVLLPLPKGACTGWMAGIPGHPGHNGAPGRDGRDGTPGEKGEKGDPGLIGPKGDIGETGVPGAEGPRGFPGIQGRKGEPGEGAYVYRSAFSVGLETYVTIPNMPIRFTKIFYNQQNHYDGSTGKFHCNIPGLYYFAYHITVYMKDVKVSLFKKDKAMLFTYDQYQENNVDQASGSVLLHLEVGDQVWLQVYGEGERNGLYADNDNDSTFTGFLLYHDTN
Source
Human pooled serum
Purity
>90%
SDS-PAGE Gel
SDS-PAGE analysis of Human Adiponectin native protein, 12% gel stained with Coomassie Brillant Blue R250
- M.W. marker – 14, 21, 31, 45, 66, 97 kDa
- reduced and heated sample, 2.5μg/lane – both bands were identified as adiponectin (LC-MS/MS)
- non-reduced and non-heated sample, 2.5μg/lane
Endotoxin
< 1.0 EU/ug
Formulation
Filtered (0,4 μm) and lyophilized in 0.5 mg/ml in 20mM TRIS, 50mM NaCl, 1mM CaCl2, pH 7.5
Reconstitution
Add deionized water to prepare a working stock solution of approximately 0.5 mg/mL and let the lyophilized pellet dissolve completely. Product is not sterile! Please filter the product by an appropriate sterile filter before using it in the cell culture.
Applications
Western blotting, ELISA, Cell culture and/or animal studies, Immunological methods
Shipping
At ambient temperature. Upon receipt, store the product at the temperature recommended below.
Storage/Expiration
Store the lyophilized protein at –80 °C. Lyophilized protein remains stable until the expiry date when stored at –80 °C. Aliquot reconstituted protein to avoid repeated freezing/thawing cycles and store at –80 °C for long term storage. Reconstituted protein can be stored at 4 °C for a week.
Quality Control Test
BCA to determine quantity of the protein.
SDS PAGE to determine purity of the protein.
Gel permeation chromatography to determine purity and oligomeric state of the native protein.
LAL to determine quantity of endotoxin.
Immunoreactivity is confirmed using monoclonal antibodies specific to human adiponectin.
Note
All blood samples used for protein preparation were tested and found negative for HBsAg, anti-HCV, HIV Ag/Ab, and syphilis. Since no test can absolutely assure the absence of all infectious agents, this product should be handled as a potential biohazard. This product is intended for research use only.
