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Catalog Number | orb669068 |
---|---|
Category | Antibodies |
Description | MitoNEET/CISD1 Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, IHC, WB |
Reactivity | Human, Monkey, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human MitoNEET/CISD1 recombinant protein (Position: Y28-E93). |
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, Monkey Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human Direct ELISA, 0.1-0.5μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 15 kDa |
UniProt ID | Q9NZ45 |
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 MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (orb669068). 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 SW620 whole cell lysates, Lane 2: monkey heart tissue lysates, Lane 3: rat heart tissue lysates, Lane 4: rat kidney tissue lysates, Lane 5: mouse heart 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 rabbit anti-MitoNEET/CISD1 antigen affinity purified polyclonal antibody (Catalog # orb669068) 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-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 (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for MitoNEET/CISD1 at approximately 15KD. The expected band size for MitoNEET/CISD1 is at 12KD.
IHC analysis of MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (orb669068). MitoNEET/CISD1 was detected in paraffin-embedded section of human gastric 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 rabbit anti-MitoNEET/CISD1 Antibody (orb669068) 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 MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (orb669068). MitoNEET/CISD1 was detected in paraffin-embedded section of human gastric 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 rabbit anti-MitoNEET/CISD1 Antibody (orb669068) 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 MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (orb669068). MitoNEET/CISD1 was detected in paraffin-embedded section of human gastric 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 rabbit anti-MitoNEET/CISD1 Antibody (orb669068) 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 MitoNEET/CISD1 using anti-MitoNEET/CISD1 antibody (orb669068). MitoNEET/CISD1 was detected in paraffin-embedded section of human rectal 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 rabbit anti-MitoNEET/CISD1 Antibody (orb669068) 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.
Flow Cytometry analysis of PC-3 cells using anti-MitoNEET/CISD1 antibody (orb669068). Overlay histogram showing PC-3 cells stained with orb669068 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MitoNEET/CISD1 Antibody (orb669068, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
ICC, IF, IHC-Fr, IHC-P | |
Bovine, Canine, Rat | |
Rabbit | |
Polyclonal | |
Unconjugated |
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