Detects human Pro-Collagen I alpha 1 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human COL3A1 is observed.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Pro Collagen I alpha1 Antibody

Detection of Human Pro-Collagen I alpha 1 by Western Blot.

Western blot shows lysates of human kidney tissue, human skin tissue, and human cartilage tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Pro-Collagen I alpha 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6220) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Pro-Collagen I alpha 1 at approximately 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human Collagen I by Western Blot.

Western blot shows lysates of IMR-90 human lung fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Pro Collagen I alpha1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6220) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). 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 Sheep Anti-Human Pro-Collagen I alpha 1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6220) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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Applications for Human Pro Collagen I alpha1 Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed IMR‑90 human lung fibroblast cell line

Immunoprecipitation

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

Simple Western

10 µg/mL
Sample: IMR-90 human lung fibroblast cell line

Western Blot

1 µg/mL
Sample: Human kidney tissue, human skin tissue, human cartilage tissue, and IMR‑90 human lung fibroblast cell line

Reviewed Applications

Read 1 review rated 5 using AF6220 in the following applications:

Immunocytochemistry/Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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

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COA

SDS

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Product Specific Notices for Human Pro Collagen I alpha1 Antibody

For research use only

Citations
Reviews

Human Pro Collagen I alpha1 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF6220

Key Product Details

Species Reactivity

Validated:

Cited:

Applications