SDHA Rabbit Polyclonal Antibody, Unconjugated

Artikelnummer: BYT-ORB654301
Artikelname: SDHA Rabbit Polyclonal Antibody, Unconjugated
Artikelnummer: BYT-ORB654301
Hersteller Artikelnummer: orb654301
Alternativnummer: BYT-ORB654301-100
Hersteller: Biorbyt
Wirt: Rabbit
Kategorie: Antikörper
Applikation: FC, ICC, IF, WB
Spezies Reaktivität: Human, Mouse, Rat
Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human SDHA, identical to the related mouse and rat sequences.
Konjugation: Unconjugated
Alternative Synonym: FP, SDH2, SDHA, SDHF
Anti-SDHA Antibody. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Klonalität: Polyclonal
Konzentration: 500 µg/ml
Molekulargewicht: 73 kDa
UniProt: P31040
Puffer: Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Formulierung: Lyophilized
Target-Kategorie: Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial
Application Verdünnung: Western blot, 0.1-0.25µg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 2µg/ml, Human Flow Cytometry (Fixed), 1-3µg/1x10 6 cells, Human, Mouse, Rat
Flow Cytometry analysis of C6 cells using anti-SDHA antibody. Overlay histogram showing C6 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-SDHA Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. 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 Hela cells using anti-SDHA antibody. Overlay histogram showing Hela 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-SDHA Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. 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 RAW264.7 cells using anti-SDHA antibody. Overlay histogram showing RAW264.7 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-SDHA Antibody (1 µg/1x10 6 cells) for 30 min at 20C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10 6 cells) was used as secondary antibody for 30 minutes at 20C. 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 SDHA using anti-SDHA antibody. SDHA was detected in immunocytochemical section of Hela cells. 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-SDHA Antibody overnight at 4C. DyLight488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Western blot analysis of SDHA using anti-SDHA 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 Hela whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human HEK293 whole cell lysates, Lane 4: human Caco-2 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat kidney tissue lysates, Lane 7: rat spleen tissue lysates, Lane 8: rat liver tissue lysates, Lane 9: mouse brain tissue lysates, Lane 10: mouse kidney tissue lysates, Lane 11: mouse liver 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-SDHA antigen affinity purified polyclonal antibody at 0.25 µg/mL overnight at 4C, 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200