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Catalog Number | orb738493 |
---|---|
Category | Antibodies |
Description | REA/PHB2 Antibody (monoclonal, 2B5) |
Species/Host | Mouse |
Clonality | Monoclonal |
Clone Number | 2B5 |
Tested applications | FC, IHC, WB |
Reactivity | Human, Mouse, Rat |
Isotype | Mouse IgG2b |
Immunogen | E.coli-derived human REA/PHB2 recombinant protein (Position: M1-K299). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.1-0.25μg/ml, Human, Mouse, Rat Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat Flow Cytometry, 1-3μg/1x106 cells, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 32 kDa |
UniProt ID | Q99623 |
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. |
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. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). 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 30ug of sample under reducing conditions. Lane 1: human HeLa whole cell lysates, Lane 2: human HEK293 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: rat heart tissue lysates, Lane 5: rat liver tissue lysates, Lane 6: mouse brain tissue lysates, Lane 7: mouse heart tissue lysates, Lane 8: mouse liver 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-REA/PHB2 antigen affinity purified monoclonal antibody (Catalog # orb738493) at 0.25 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for REA/PHB2 at approximately 32KD. The expected band size for REA/PHB2 is at 32KD.
IHC analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). REA/PHB2 was detected in paraffin-embedded section of human gallbladder adenocarcinoma 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 2μg/ml mouse anti-REA/PHB2 Antibody (orb738493) 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) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). REA/PHB2 was detected in paraffin-embedded section of human ovarian serous adenocarcinoma 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 2μg/ml mouse anti-REA/PHB2 Antibody (orb738493) 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) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). REA/PHB2 was detected in paraffin-embedded section of human renal clear cell carcinoma 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 2μg/ml mouse anti-REA/PHB2 Antibody (orb738493) 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) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). REA/PHB2 was detected in paraffin-embedded section of human lung 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 2μg/ml mouse anti-REA/PHB2 Antibody (orb738493) 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) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). REA/PHB2 was detected in paraffin-embedded section of mouse colon 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 2μg/ml mouse anti-REA/PHB2 Antibody (orb738493) 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) (Catalog # orb90443) with DAB as the chromogen.
IHC analysis of REA/PHB2 using anti-REA/PHB2 antibody (orb738493). REA/PHB2 was detected in paraffin-embedded section of rat colon 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 2μg/ml mouse anti-REA/PHB2 Antibody (orb738493) 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) (Catalog # orb90443) with DAB as the chromogen.
Flow Cytometry analysis of HepG2 cells using anti-REA/PHB2 antibody (orb738493). Overlay histogram showing HepG2 cells stained with orb738493 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-REA/PHB2 Antibody (orb738493, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. 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.
FC, IHC, WB | |
Human, Mouse, Rat | |
Mouse | |
Monoclonal | |
Unconjugated |
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