Cyclophilin A/PPIA Antibody

SKU: orb19174

Description

Cyclophilin A/PPIA Antibody

Images & Validation

• Tested Applications: FC, ICC, IF, IHC, IHC-Fr, WB
• Reactivity: Human, Mouse, Rat
• Application Notes:
  Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat Western blot, 0.1-0.5μg/ml, Human, Mouse, RatImmunohistochemistry (Frozen Section), 0.5-1μg/ml, Human Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human Cyclophilin A recombinant protein (Position: T116-E165). Human Cyclophilin A shares 98% and 95.9% amino acid (aa) sequence identity with mouse and rat Cyclophilin A, respectively.
• Molecular Weight: 185 kDa
• Purification: Immunogen affinity purified.

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
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Disclaimer: For research use only

Alternative Names

Peptidyl-prolyl cis-trans isomerase A; PPIase A; 5.2.1.8; Cyclophilin A; Cyclosporin A-binding protein; Rotamase A; Peptidyl-prolyl cis-trans isomerase A, N-terminally processed; PPIA; CYPA

Flow Cytometry analysis of K562 cells using anti-PPIA antibody. Overlay histogram showing K562 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPIA Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of THP-1 cells using anti-PPIA antibody. Overlay histogram showing THP-1 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPIA Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of U937 cells using anti-PPIA antibody. Overlay histogram showing U937 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPIA Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in immunocytochemical section of A549 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL rabbit anti-Cyclophilin A Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit 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.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in paraffin-embedded section of human intestinal cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Cyclophilin A Antibody 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) with DAB as the chromogen.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Cyclophilin A Antibody 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) with DAB as the chromogen.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in paraffin-embedded section of rat spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Cyclophilin A Antibody 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) with DAB as the chromogen.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in immunocytochemical section of THP-1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/ml rabbit anti-Cyclophilin A Antibody overnight at 4°C. Biotin conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in immunocytochemical section of THP-1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/ml rabbit anti-Cyclophilin A Antibody overnight at 4°C. Biotin conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in immunocytochemical section of THP-1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/ml rabbit anti-Cyclophilin A Antibody overnight at 4°C. Biotin conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Cyclophilin A using anti-Cyclophilin A antibody. Cyclophilin A was detected in immunocytochemical section of THP-1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/ml rabbit anti-Cyclophilin A Antibody overnight at 4°C. Biotin conjugated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using DyLight®488 Conjugated Avidin. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of Cyclophilin A using anti-Cyclophilin A 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 30 ug of sample under reducing conditions. Lane 1: human THP-1 whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human MOLT-4 whole cell lysates, Lane 4: human Daudi whole cell lysates, Lane 5: rat lung tissue lysates, Lane 6: rat brain tissue lysates, Lane 7: mouse lung tissue lysates, Lane 8: mouse brain 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-Cyclophilin A antigen affinity purified polyclonal 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-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 with Tanon 5200 system. A specific band was detected for Cyclophilin A at approximately 185 kDa. The expected band size for Cyclophilin A is at 185 kDa.

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UniProt

P62937