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Catalog Number | orb865599 |
---|---|
Category | Antibodies |
Description | Acid phosphatase/ACP5 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human Acid phosphatase/ACP5 recombinant protein (Position: N39-P325). |
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, Human, Mouse, Rat Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human, Mouse Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry, 1-3 μg/1x106 cells, Human Direct ELISA, 0.1-0.5 μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 39 kDa |
UniProt ID | P13686 |
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 Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (orb865599). 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 HepG2 whole cell lysates, Lane 2: human A431 whole cell lysates, Lane 3: human Jurkat whole cell lysates, Lane 4: human HeLa whole cell lysates, Lane 5: human K562 whole cell lysates, Lane 6: human Raji whole cell lysates, Lane 7: rat lung tissue lysates, Lane 8: mouse lung 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-Acid Phosphatase/ACP5 antigen affinity purified polyclonal antibody (Catalog # orb865599) 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 Acid Phosphatase/ACP5 at approximately 39 kDa. The expected band size for Acid Phosphatase/ACP5 is at 37 kDa.
IHC analysis of Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (orb865599). Acid Phosphatase/ACP5 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-Acid Phosphatase/ACP5 Antibody (orb865599) 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 Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (orb865599). Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of mouse lung 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-Acid Phosphatase/ACP5 Antibody (orb865599) 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 Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (orb865599). Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of human lung 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-Acid Phosphatase/ACP5 Antibody (orb865599) 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 Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (orb865599). Acid Phosphatase/ACP5 was detected in a paraffin-embedded section of human renal cancer 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-Acid Phosphatase/ACP5 Antibody (orb865599) 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 Acid Phosphatase/ACP5 using anti-Acid Phosphatase/ACP5 antibody (orb865599). Acid Phosphatase/ACP5 was detected in an 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 5 μg/mL rabbit anti-Acid Phosphatase/ACP5 Antibody (orb865599) 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.
Flow Cytometry analysis of U251 cells using anti-Acid Phosphatase/ACP5 antibody (orb865599). Overlay histogram showing U251 cells stained with orb865599 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Acid Phosphatase/ACP5 Antibody (orb865599, 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 (Red line) was also used as a control.
FC, IF, IHC | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
FC, IF, IHC | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
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