DHFR Mouse Monoclonal Antibody

SKU: orb527060

Description

DHFR Mouse Monoclonal Antibody

Research Area

Cancer Biology, Cell Biology, Metabolism Research, Protein Biochemistry, Signal Transduction

Images & Validation

• Tested Applications: IHC, WB
• Dilution Range: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
• Reactivity: Human, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Mouse
• Clonality: Monoclonal
• Isotype: Mouse IgG2b
• Clone No.: 3C8
• Immunogen: E.coli-derived human DHFR recombinant protein (Position: V2-D187). Human DHFRshares90%aminoacid (aa)sequenceidentitywith both mouseandrat DHFR.
• Target: Dihydrofolate reductase
• Molecular Weight: 22 kDa
• Purification: Immunogen affinity purified.
• 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: Lyophilized
• Buffer/Preservatives: Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

DHFR; DHFRP1; dihydrofolate reductase

IHC analysis of DHFR using anti-DHFR antibody. DHFR was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-DHFR Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of DHFR using anti-DHFR antibody. DHFR was detected in paraffin-embedded section of rat gaster tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-DHFR Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of DHFR using anti-DHFR antibody. 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 50 ug of sample under reducing conditions. Lane 1: human K562 whole cell lysates, Lane 2: human SGC-7901 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human SW620 whole cell lysates, Lane 5: rat kidney tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-DHFR antigen affinity purified monoclonal antibody 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for DHFR at approximately 22KD. The expected band size for DHFR is at 22KD.

Quick Database Links

Gene Symbol

Dihydrofolate reductase

UniProt

P00374