Western blot shows lysates of IMR-90 human lung fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human N-Pro Collagen I Monoclonal Antibody (Catalog # MAB6220) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Collagen I at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Pro-Collagen I alpha 1 in IMR‑90 Human Cell Line.

Pro-Collagen I alpha 1 was detected in immersion fixed IMR-90 human lung fibroblast cell line using Mouse Anti-Human Pro-Collagen I alpha 1 Monoclonal Antibody (Catalog # MAB6220) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Pro-Collagen I alpha 1 by Immunoprecipitation.

Human Pro-Collagen I alpha 1 was immunoprecipitated from 500 μg of IMR-90 human lung fibroblast cell line lysates with 5 μg of Mouse Anti-Human Pro-Collagen I alpha 1 Monoclonal Antibody (Catalog # MAB6220). The N-Pro Collagen I-antibody complexes were absorbed using Protein A or Protein G. Immunoprecipitated human Pro-Collagen I alpha 1 was detected by Western blot using 1 μg/mL of Sheep Anti-Human Pro-Collagen I alpha 1 Antigen Affinity-purified Polyclonal Antibody (AF6220) under reducing conditions and using Western Blot Buffer Group 1.

Applications for Human Pro Collagen I alpha1 Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed IMR-90 human lung fibroblast cell line

Immunoprecipitation

5 µg/mL
Sample: IMR‑90 human lung fibroblast cell line

Western Blot

1 µg/mL
Sample: IMR‑90 human lung fibroblast cell line

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Collagen I alpha 1

Type I collagen is the most abundant structural protein of connective tissues such as skin, bone and tendon. It is synthesized as a procollagen molecule which is characterized by a 300 nm triple helical domain flanked by globular N- and C-terminal propeptides (1). The triple helical domain contains Gly-Xaa-Yaa triplets where Xaa and Yaa are frequently proline and hydroxyproline, respectively. The non-helical propeptides are removed by procollagen N- and C-proteinase activities so that the mature triple helices can self-assemble into collagen fibrils that provide tensile strength to tissues (1). Type I collagen is a heterotrimer that consists of two alpha1(I) chains and one alpha2(I) chain, although homotrimers consisting of three identical alpha1(I) chains have also been described (2). This recombinant mini pro-alpha 1(I) collagen consists of a shortened alpha1(I) chain with following domain structure from N- to C-terminus: N-propeptide, N‑telopeptide, the 33 most N-terminal Gly-Xaa-Yaa repeats, the 33 most C-terminal Gly-Xaa-Yaa repeats, C-telopeptide and C-propeptide. The preparation contains a mixture of the full-length molecule, pN collagen I( alpha1) and the C-terminal propeptide. This truncated pro-alpha 1(I) collagen is a substrate for procollagen N-proteinase and procollagen C-proteinase.

References

Canty, E.G. et al. (2005) J. Cell Sci. 118:1341.
Han, S. et al. (2008) J. Mol. Biol. 383:122.

Alternate Names

COL1A1, OI4

Entrez Gene IDs

1277 (Human); 12842 (Mouse); 29393 (Rat)

Gene Symbol

COL1A1

UniProt

P02452

Additional Collagen I alpha 1 Products

Product Documents for Human Pro Collagen I alpha1 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human Pro Collagen I alpha1 Antibody

For research use only

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