Recombinant Human Pro-Collagen III alpha 1/COL3A1, CF

Catalog # Availability Size / Price Qty
7294-CL-020
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Recombinant Human Pro-Collagen III alpha 1/COL3A1, CF Summary

Product Specifications

Purity
>75%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
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
Accession #
N-terminal Sequence
Analysis
His
Predicted Molecular Mass
40 kDa
SDS-PAGE
41-46 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

7294-CL

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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • 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
  1. Dilute rhPro-COL3A1 to 75 μg/mL in Assay Buffer.
  2. Dilute rhBMP-1 to 5 μg/mL in Assay Buffer.
  3. 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.
  4. Incubate reaction vials and controls at 37 °C for one hour.
  5. 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.
  6. 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.
  7. 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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

References
  1. Kuivaniemi H. et al. (1997) Hum. Mutat. 9:300.
Entrez Gene IDs
1281 (Human); 12825 (Mouse); 84032 (Rat)
Alternate Names
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

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