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Collagen I Rabbit Polyclonal Antibody
Description
Research Area
Images & Validation
−| Tested Applications | IF, IHC-Fr, IHC-P, WB |
|---|---|
| Dilution Range | WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500 |
| Reactivity | Human, Mouse, Rabbit, Rat |
| Predicted Reactivity | Bovine, Canine, Gallus, Sheep |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | KLH conjugated synthetic peptide derived from human Collagen I C-terminal propeptide (1321-1400/1464aa) |
| Target | COL1A1 |
| Molecular Weight | 130 kDa |
| Purification | affinity purified by Protein A |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles. |
|---|---|
| Form/Appearance | Liquid |
| Buffer/Preservatives | 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol. |
| Concentration | 1mg/ml |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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Blank control: Hela. Primary Antibody (green line): Rabbit Anti-Collagen I antibody (orb312178), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with Collagen I Polyclonal Antibody, Unconjugated (orb312178) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum, Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Antibody incubation with Collagen I Polyclonal Antibody, Unconjugated (orb312178) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit) and DAB staining.

Sample: Lane 1: HepG2 (Human) Cell Lysate at 30 ug, Primary: Anti-Collagen I (orb312178) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 130, 110 kD, Observed band size: 130 kD.

Sample: Lane 1: Placenta (Mouse) Lysate at 40 ug, Lane 2: U-2OS (Human) Cell Lysate at 30 ug, Primary: Anti-Collagen I (orb312178) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 130 kD, Observed band size: 130 kD.

Sample: Lane 1: Placenta (Mouse) Lysate at 40 ug, Lane 2: U-2OS (Human) Cell Lysate at 30 ug, Primary: Anti-Collagen I (orb312178) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 130, 110 kD, Observed band size: 130 kD.

Sample: Ovary (Mouse) Lysate at 40 ug, NIH/3T3 (Mouse) Cell Lysate at 40 ug, Liver (Rat) Lysate at 40 ug, 293T (huamn) Cell Lysate at 40 ug, A431 (huamn) Cell Lysate at 40 ug, RAW264.7 (Mouse) Cell Lysate at 40 ug, Primary: Anti-Collagen I (orb312178) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 130 kD, Observed band size: 110 kD.

Tissue/Cell: human lung carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-Collagen I Polyclonal Antibody, Unconjugated (orb312178) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
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Syverud, Brian C. et al. Quantitative, Label-Free Evaluation of Tissue-Engineered Skeletal Muscle Through Multiphoton Microscopy Tissue Eng Part C Methods, 23, 616-626 (2017)
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Collagen I Rabbit Polyclonal Antibody (orb312178)
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