- R1038 - OriGene COL1A1 antibody

Villone D, Fritsch A, Koch M, Bruckner-Tuderman L, Hansen U, Bruckner P
The Journal of biological chemistry 2008 Sep 5;283(36):24506-13

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Supportive validation

Submitted by: OriGene (provider)
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Experimental details: Western Blot of Human Collagen Type?I using Collagen I Antibody .? Lane 1: Human Collagen Type 1. Lane 2: None. Load: 50 ng per lane. Primary antibody: Collagen Type I antibody at 1:1,000 overnight at 4°C. Secondary antibody: DyLight?TM 649 rabbit secondary antibody at 1:20,000 for 30 min at RT. Block: MB-070 for 30 min at RT. Predicted/Observed size: 139 & 130 kDa, 139 & 130 kDa for Collagen Type I. Other Band(s): Collagen Type I splice variants and isoforms.
Validation comment: WB

Supportive validation

Submitted by: OriGene (provider)
Main image
Experimental details: Western blot analysis is shown using Collagen type I antibody to detect expression of collagen I in Wistar rat hepatic stellate cells (HSC) in control (GFP-transduced) (left lane) and PPARgamma-transduced cell lysates (right lane). Protein staining shown below each blot depicts equal protein loading. An equal amount of the whole cell protein (100 ug) was separated by SDS-PAGE and electroblotted to nitrocellulose membranes. Proteins were detected by incubating the membrane with Collagen type I antibody at a concentration of 0.2-2 ug/10 ml in TBS (100 mM Tris-HCl, 0.15 M NaCl, pH 7.4) with 5% Non-fat milk. Detection occurred by incubation with a horseradish peroxidase-conjugated secondary antibody at 1 ug/10 ml. Proteins were detected by a chemiluminescent method using the PIERCE ECL kit (Amersham Biosciences). Other detection systems will yield similar results. See Hazra et al. (2004) for additional details.
Validation comment: WB

Supportive validation

Submitted by: OriGene (provider)
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Experimental details: Immunohistochemistry using affinity purified Collagen type I antibody at a 1/100 dilution to detect distal tubules in normal kidney tissue. Note the absence of staining of glomeruli. The antibody was reacted with antibody for 4 h RT followed by secondary antibody and substrate reaction. Tissue was Formalin-fixed and Paraffin embedded. No antigen retrieval was performed.
Validation comment: IHC

Supportive validation

Submitted by: OriGene (provider)
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Experimental details: Immunohistochemistry of human lung tissue (Formalin-fixed, Paraffin-embedded) using Collagen type I antibody : Primary antibody (Collagen I) at 1:400, secondary antibody: Peroxidase goat anti-rabbit at 1:10,000 for 45 min at RT; Localization: Strong staining was observed in the extracellular matrix of the lung. Epithelial cells were negative; Staining: Antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.
Validation comment: IHC

Supportive validation

Submitted by: OriGene (provider)
Main image
Experimental details: Immunohistochemistry of a liver section (Formalin-fixed, Paraffin-embedded) using Collagen type I antibody .? A: Central vein (CV) fibrosis, B: Non-fibrotic CV, C: Perisinusodial fibrosis, D: Non-fibrotic area, E: Protat tract fibrosis, F: Septal fibrosis (arrow). Primary antibody: Collagen type I antibody at 1:1250 for 4°C for 24hr; Secondary antibody: Peroxidase biotin-streptavidin rabbit secondary antibody at 1:10,000 for 45 min at RT; Localization: Collagen type I is intra- and extracellular; Staining: 3.3-diaminobenzidine tetrahydrochloride was used as the chromogen. Nuclei were counterstained purple with hematoxylin.
Validation comment: IHC

R1038