MPI Antibody (monoclonal, 11I4)

• Catalog Number: orb654265
• Category: Antibodies
• Description: MPI Antibody (monoclonal, 11I4)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 11I4
• Tested applications: FC, ICC, IF, WB
• Reactivity: Human
• Isotype: Mouse IgG1
• Immunogen: E. coli-derived human MPI recombinant protein (Position: A2-K99). Human MPI shares 88.8% and 86.7% amino acid (aa) sequence identity with mouse and rat MPI, respectively.
• Concentration: 0
• Dilution range: Western blot, 0.1-0.5μg/ml, Human Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 47 kDa
• UniProt ID: P34949
• Sensitivity: > 5000 cells
• Storage: Store 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 freeze-thaw cycles.
• Alternative names: Mannose-6-phosphate isomerase; Phosphohexomutase;
• Note: For research use only
• Application notes: Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of MPI using anti-MPI antibody (orb654265). 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 50ug of sample under reducing conditions. Lane 1: human Caco-2 whole cell lysates Lane 2: human HeLa whole cell 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-MPI antigen affinity purified monoclonal antibody (Catalog # orb654265) at 0.5 μg/mL overnight at 4℃, 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for MPI at approximately 47KD. The expected band size for MPI is at 47KD.

Flow Cytometry analysis of SiHa cells using anti-MPI antibody (orb654265). Overlay histogram showing U20S cells stained with orb654265 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-MPI Antibody (orb654265, 1μg/1x10^6 cells) for 30 min at 20℃. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20℃. Isotype control antibody (Green line) was mouse IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

IF analysis of MPI using anti-MPI antibody (orb654265). MPI was detected in immunocytochemical section of MCF-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 5μg/mL mouse anti-MPI Antibody (orb654265) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 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.