Collagen I/COL1A1 Antibody

• Catalog Number: orb654259
• Category: Antibodies
• Description: Collagen I/COL1A1 Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IHC, WB
• Predicted Reactivity: Hamster
• Reactivity: Human, Mouse
• Isotype: Rabbit IgG
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Collagen I (1203-1218aa QPPQEKAHDGGRYYRA), different from the related rat and mouse sequences by two amino acids.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5μg/ml, Human, Mouse Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, By Heat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 138941 MW
• UniProt ID: P02452
• Storage: At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
• Alternative names: Collagen alpha-1 (I) chain;Alpha-1 type I collagen
• Note: For research use only
• Application notes: WB: The detection limit for COL1A1 is approximately 0.5ng/lane under reducing conditions. Tested Species: In-house tested species with positive results. Predicted Species: Species predicted to be fit for the product based on sequence similarities. By Heat: Boiling the paraffin sections in 10mM citrate buffer, pH6.0, for 20mins is required for the staining of formalin/paraffin sections. Other applications have not been tested. Optimal dilutions should be determined by end users. . Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of COL1A1 using anti-COL1A1 antibody (orb654259). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel)/90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human HK2 whole cell lysates, Lane 2: human HCC tissue lysates, Lane 3: mouse NIH/3T3 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-COL1A1 antigen affinity purified polyclonal antibody (Catalog # orb654259) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) ?with Tanon 5200 system. A specific band was detected for COL1A1 at approximately 180-200 kDa. The expected band size for COL1A1 is at 138 kDa.

IHC analysis of COL1A1 using anti-COL1A1 antibody (orb654259). COL1A1 was detected in a paraffin-embedded section of human colon adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-COL1A1 Antibody (orb654259) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.

IHC analysis of COL1A1 using anti-COL1A1 antibody (orb654259). COL1A1 was detected in a paraffin-embedded section of human gastric carcinomaa tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-COL1A1 Antibody (orb654259) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.

IHC analysis of COL1A1 using anti-COL1A1 antibody (orb654259). COL1A1 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-COL1A1 Antibody (orb654259) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.