Recombinant Human Pro-Collagen III alpha 1/COL3A1, CF
Recombinant Human Pro-Collagen III alpha 1/COL3A1, CF Summary
Thr1097-Leu1466, with an N-terminal 6-His tag
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.|
|Shipping||The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- Assay Buffer: 25 mM HEPES, 0.01% Brij, pH 7.5
- Recombinant Human Pro Collagen III alpha 1/COL3A1 (rhPro-COL3A1) (Catalog # 7294-CL)
- Recombinant Human BMP‑1/PCP (rhBMP-1) (Catalog # 1927-ZN)
- Reducing SDS-PAGE Sample Buffer
- SDS-PAGE or Western Blot
- Dilute rhPro-COL3A1 to 75 μg/mL in Assay Buffer.
- Dilute rhBMP-1 to 5 μg/mL in Assay Buffer.
- Combine one volume of diluted rhPro-COL3A1 with three volumes of diluted rhBMP-1. For controls, combine one volume of rhPro‑COL3A1 with three volumes of Assay Buffer, as well as three volumes of rhBMP-1 with one volume of Assay Buffer.
- Incubate reaction vials and controls at 37 °C for one hour.
- After incubation, combine rhPro-COL3A1/rhBMP-1 reaction mixtures and controls with reducing SDS-PAGE gel buffer at a 2:1 (reaction mixture:gel buffer) ratio (v/v). Mix and incubate samples at 95-100 °C for 3-5 minutes to stop reactions.
- Load 40 μL (0.5 μg of rhPro-COL3A1) per lane and analyze the cleavage by SDS-PAGE followed by protein staining and/or Western blot.
- Activity calculation:
|% Cleavage = [1 -||% full-length rhPro-COL3A1 (reaction)||] x 100%|
|% full-length rhPro-COL3A1 (control)|
- rhCOL3A1: 0.5 µg
- rhBMP-1: 0.1 µg
Background: Collagen III alpha 1/COL3A1
Collagen type III is found in most soft connective tissues, such as skin, lung, and the vascular system, often in association with type I collagen. It is a trimer of
alpha 1(III) chains that are linked by interchain disulfide bonds. Defective collagen type III is a cause of Ehlers-Danlos syndrome types III and IV and arterial aneurisms (1). This recombinant mini pro-alpha 1(III) collagen consists of a triple-helical region of Gly-Xaa-Yaa repeats, a short non-helical region, and the C‑terminal propeptide. The C‑terminal propeptide can be removed by the procollagen C‑proteinase (BMP-1).
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