Recombinant Human Pro-Collagen III alpha 1/COL3A1, CF Summary
Details of Functionality
Measured by the cleavage of its C-terminal propeptide by Recombinant Human BMP‑1/PCP (Catalog # 1927-ZN). >50% of full-length Pro-Collagen III alpha 1 is cleaved by recombinant human BMP-1, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human Collagen III alpha 1/COL3A1 protein Thr1097-Leu1466, with an N-terminal 6-His tag
>75%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.
Applications/Dilutions
Dilutions
Enzyme Activity
Theoretical MW
40 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.
SDS-PAGE
41-46 kDa, reducing conditions
Publications
Read Publications using 7294-CL in the following applications:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
6 months from date of receipt, -20 to -70 °C as supplied.
3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Sodium Acetate and NaCl.
Purity
>75%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Assay Procedure
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)
Per Lane:
rhCOL3A1: 0.5 µg
rhBMP-1: 0.1 µg
Notes
Coomassie is a registered trademark of Imperial Chemical Industries Ltd.
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human Pro-Collagen III alpha 1/COL3A1, CF
alpha1 (III) collagen
alpha-1 type III collagen
COL3A1
Collagen 3
collagen alpha-1(III) chain
Collagen III alpha 1
collagen, fetal
collagen, type III, alpha 1
Collagen-3
collagenIII
Collagen-III
EDS4A
EDSVASC
Ehlers-Danlos syndrome type IV, autosomal dominant
FLJ34534
PMGEDSV
Background
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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