Human Pro Collagen I alpha 1 Antibody (816161)

R&D Systems | Catalog # MAB6220

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Western Blot, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 816161
View all Collagen I alpha 1 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Collagen I
Gln23-Lys277, Gly1094-Leu1464
Accession # P02452

Specificity

Detects human Pro-Collagen I alpha 1 in direct ELISAs.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Pro Collagen I alpha 1 Antibody (816161)

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 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.
Collagen I a1 antibody in IMR-90 Human Cell Line by Immunocytochemistry (ICC).

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.
Detection of gnzwt4vp4jf4h7neevex.jpg by Western Blot

Detection of gnzwt4vp4jf4h7neevex.jpg by Western Blot

SfA induces secretion of PPIB from cells.(A) Western blot of intracellular and extracellular PPIB in Jurkat cells treated with 1 μM SfA or 1 μM CsA for 4 hours ± 10.6 μM brefeldin A. (B) Immunofluorescence imaging of PPIB and PDI in HeLa cells treated with 1 μM SfA or 1 μM CsA for 4 hours. Scale bars: 20 μm. (C) Volcano plot of proteins secreted by Jurkat cells following treatment with 1 μM SfA for 4 hours relative to vehicle (n = 3). (D) Volcano plot of proteins secreted by Jurkat cells following treatment with 1 μM CsA for 4 hours relative to vehicle (n = 3). (E) Structure of sanglifehrin macrocycle (SfA-mc). (F) Western blot of intracellular PPIB in Jurkat cells treated with 1 μM SfA or 1 μM SfA-mc for 4 hours ± 10.6 μM brefeldin A. (G) Western blot of Jurkat cell lysates for ER stress markers after treatment with 1 μM SfA or 1 μM SfA-mc with or without brefeldin A for 24 hours (n = 3). Image collected and cropped by CiteAb from the following open publication (https://insight.jci.org/articles/view/171162), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of il7gn6ioyvz0zygdjdsk.jpg by Western Blot

Detection of il7gn6ioyvz0zygdjdsk.jpg by Western Blot

SfA reduces collagen secretion from primary fibrotic fibroblasts.(A and B) Generation of precision cut lung slices (PCLS) from explanted lung tissue isolated from patients with idiopathic pulmonary fibrosis (IPF). Lung tissue was obtained in 1 cm blocks (A) before being sliced (200–300 μm thick) using a Compresstome VF-310-0Z (B). (C) PCLS were then treated with or without SfA (1 μM) for 72 hours in culture (n = 4). (D–G) Analysis of collagen and PPIB secretion from PCLS prepared from IPF patient lung tissue ± 1 μM SfA in supernatants (n = 4). (H) Western blot for collagen type I secreted by primary human fibroblasts isolated from healthy controls and patients with IPF ± 1 μM SfA over 96 hours in cell supernatants. (I) Intracellular PPIB levels in healthy or IPF fibroblasts with or without 1 μM SfA treatment by immunofluorescence. (J) Western blot for alpha -smooth muscle actin ( alpha SMA) and phosphoSMAD3/SMAD3 signaling in IPF fibroblasts ± SfA over 96 hours in cell lysates. GAPDH was used as loading control. (K) COL1A1 levels in cell lysates and supernatants of TGF-beta –activated fibroblasts treated with or without 1 μM SfA and 10 μM brefeldin A. D–G were analyzed with a 2-tailed t test against a hypothetical value of 1. Graphed data in I and K represent mean ± SD. Image collected and cropped by CiteAb from the following open publication (https://insight.jci.org/articles/view/171162), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Pro-Collagen I alpha 1 by Western Blot

Detection of Pro-Collagen I alpha 1 by Western Blot

SfA induces secretion of PPIB from cells.(A) Western blot of intracellular and extracellular PPIB in Jurkat cells treated with 1 μM SfA or 1 μM CsA for 4 hours ± 10.6 μM brefeldin A. (B) Immunofluorescence imaging of PPIB and PDI in HeLa cells treated with 1 μM SfA or 1 μM CsA for 4 hours. Scale bars: 20 μm. (C) Volcano plot of proteins secreted by Jurkat cells following treatment with 1 μM SfA for 4 hours relative to vehicle (n = 3). (D) Volcano plot of proteins secreted by Jurkat cells following treatment with 1 μM CsA for 4 hours relative to vehicle (n = 3). (E) Structure of sanglifehrin macrocycle (SfA-mc). (F) Western blot of intracellular PPIB in Jurkat cells treated with 1 μM SfA or 1 μM SfA-mc for 4 hours ± 10.6 μM brefeldin A. (G) Western blot of Jurkat cell lysates for ER stress markers after treatment with 1 μM SfA or 1 μM SfA-mc with or without brefeldin A for 24 hours (n = 3). Image collected and cropped by CiteAb from the following open publication (https://insight.jci.org/articles/view/171162), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Pro-Collagen I alpha 1 by Western Blot

Detection of Pro-Collagen I alpha 1 by Western Blot

SfA induces secretion of PPIB from cells.(A) Western blot of intracellular and extracellular PPIB in Jurkat cells treated with 1 μM SfA or 1 μM CsA for 4 hours ± 10.6 μM brefeldin A. (B) Immunofluorescence imaging of PPIB and PDI in HeLa cells treated with 1 μM SfA or 1 μM CsA for 4 hours. Scale bars: 20 μm. (C) Volcano plot of proteins secreted by Jurkat cells following treatment with 1 μM SfA for 4 hours relative to vehicle (n = 3). (D) Volcano plot of proteins secreted by Jurkat cells following treatment with 1 μM CsA for 4 hours relative to vehicle (n = 3). (E) Structure of sanglifehrin macrocycle (SfA-mc). (F) Western blot of intracellular PPIB in Jurkat cells treated with 1 μM SfA or 1 μM SfA-mc for 4 hours ± 10.6 μM brefeldin A. (G) Western blot of Jurkat cell lysates for ER stress markers after treatment with 1 μM SfA or 1 μM SfA-mc with or without brefeldin A for 24 hours (n = 3). Image collected and cropped by CiteAb from the following open publication (https://insight.jci.org/articles/view/171162), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Pro Collagen I alpha 1 Antibody (816161)

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.<br/>*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.

Calculators

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 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 alpha 1(I) chains and one alpha 2(I) chain, although homotrimers consisting of three identical alpha 1(I) chains have also been described (2). This recombinant mini pro-alpha 1(I) collagen consists of a shortened alpha 1(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( alpha 1) and the C-terminal propeptide. This truncated pro-alpha 1(I) collagen is a substrate for procollagen N-proteinase and procollagen C-proteinase.

References

  1. Canty, E.G. et al. (2005) J. Cell Sci. 118:1341.
  2. 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 alpha 1 Antibody (816161)

Certificate of Analysis

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Product Specific Notices for Human Pro Collagen I alpha 1 Antibody (816161)

For research use only

Citations for Human Pro Collagen I alpha 1 Antibody (816161)

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