CD45/Ptprc Antibody

• Catalog Number: orb865646
• Category: Antibodies
• Description: CD45/Ptprc Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IF, IHC, WB
• Reactivity: Mouse, Rat
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived mouse CD45/Ptprc recombinant protein (Position: N209-E1269).
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5 μg/ml, Mouse Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Mouse Direct ELISA, 0.1-0.5 μg/ml, Mouse
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 250 kDa
• UniProt ID: P06800
• 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.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of CD45/Ptprc using anti-CD45/Ptprc antibody (orb865646). 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: mouse spleen tissue lysates, Lane 2: mouse thymus tissue 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-CD45/Ptprc antigen affinity purified polyclonal antibody (Catalog # orb865646) 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 CD45/Ptprc at approximately 250 kDa. The expected band size for CD45/Ptprc is at 145 kDa.

IHC analysis of CD45/Ptprc using anti-CD45/Ptprc antibody (orb865646). CD45/Ptprc was detected in a paraffin-embedded section of rat lymph node 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-CD45/Ptprc Antibody (orb865646) 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 CD45/Ptprc using anti-CD45/Ptprc antibody (orb865646). CD45/Ptprc was detected in a paraffin-embedded section of rat 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-CD45/Ptprc Antibody (orb865646) 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 CD45/Ptprc using anti-CD45/Ptprc antibody (orb865646). CD45/Ptprc was detected in a paraffin-embedded section of mouse lymph node 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-CD45/Ptprc Antibody (orb865646) 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.

IF analysis of CD45/Ptprc using anti-CD45/Ptprc antibody (orb865646). CD45/Ptprc was detected in an immunocytochemical section of RAW264.7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 μg/mL rabbit anti-CD45/Ptprc Antibody (orb865646) overnight at 4°C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1: 500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.