Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7]
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ab290021 is the same product as ab110322.
View Alternative Names
AAC1, ANT1, SLC25A4, ADP/ATP translocase 1, Adenine nucleotide translocator 1, Solute carrier family 25 member 4, ANT 1, AAC2, ANT2, SLC25A5, ADP/ATP translocase 2, Adenine nucleotide translocator 2, Solute carrier family 25 member 5, ANT 2
- ICC
Supplier Data
Immunocytochemistry - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (AB290021)
Immunocytochemistry - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7].
ab290021 at 10μg/ml staining Adenine Nucleotide Translocase 1/ANT 1 in cultured Human fibroblasts (fixed and permeabilized) by Immunocytochemistry, followed by a fluorescent goat-anti-mouse IgG.
- WB
Supplier Data
Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (AB290021)
Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7].
All lanes : ab290021 at 1 μg/ml
Lane 1 : Human heart mitochondria isolate at 5 μg
Lane 2 : Bovine heart mitochondria isolate at 1 μg
Lane 3 : Rat heart mitochondria isolate at 10 μg
Lane 4 : Mouse heart mitochondria isolate at 10 μg
Predicted band size : 33 kDa
Extra bands in the mouse sample (lane 4) are due to the reaction of the IgG-specific goat anti-mouse secondary antibody with residual mouse blood in the heart tissue, as it is very difficult to entirely remove the blood from these small organs.
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- WB
Supplier Data
Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (AB290021)
Western Blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7].
All lanes : ab290021 at 1 μg/ml
Lane 1 : SLC25A4 (Human) Recombinant Protein (ANT1) cell lysate at 0.1 μg
Lane 2 : Recombinant Human Adenine Nucleotide Translocator 2/ANT 2 protein (ab157891) cell lysate at 0.1 μg
Lane 3 : Human Skeletal Muscle cell lysate at 20 μg
Lane 4 : Human Liver cell lysate at 20 μg
Lane 5 : Mouse Skeletal Muscle cell lysate at 20 μg
Lane 6 : Mouse Liver cell lysate at 20 μg
Lane 7 : Rat Skeletal Muscle cell lysate at 20 μg
Lane 8 : Rat Liver cell lysate at 20 μg
Predicted band size : 33 kDa
Observed band size : 33 kDa
False colour image of Western blot : Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] staining at 1 μg/ml, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab290021 was shown to bind specifically to Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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Reactivity data
Product details
ab290021 is the same product as ab110322. All associated reference to ab110322 apply to ab290021.
This antibody can also recognise ADP/ATP translocase 2, SLC25A31, SLC25A5 and SLC25A6 due to homology of 93% in immunogen sequence.
ab290021 was produced in vitro using hybridomas grown in serum-free medium, and then purified by biochemical fractionation.
This antibody comes with an Incubation Solution which should be used as the diluent for Western blot (the antibody should not be diluted in milk for incubations). The lab has found that diluting the antibody in this solution gives optimal results. Dilute the antibody to 1 ug/mL in neat Incubation Solution before use.
The following is a quick reference procedure guide for Western Blot, specific for ab290021:
1. Solubilize samples in SDS-PAGE sample buffer.
2. Heat sample to 37°C for 30 minutes.
3. Separate proteins on Tris-glycine 10-22% gradient gel 150 V ~ 2 hrs.
4. Soak gel in 50 mL CAPS buffer for 30 minutes.
5. Transfer to 0.45 μm PVDF membrane in CAPS buffer for 2 hrs at 0.15 A CAPS buffer : 10 mM 3-[cyclohexylamino]-1-propane sulfonic acid, pH 11 w/NaOH, 10% methanol ice cold throughout.
6. Block membrane for 3 hrs - overnight in 5% milk/PBS 4°C.
7. Wash blot 3 times for 5 mins PBS/0.05% Tween-20 with agitation. Followed by a final rinse in PBS.
8. Antibody diluted to concentration specified in supplied incubation solution for 2 hrs.
9. Wash blot 3 times in PBS/0.05% Tween-20 for 5 mins.
10. Incubate blot with secondary anti-mouse antibody diluted according to manufacturer's instructions in 1% milk/PBS for 2 hrs.
11. Wash blot 3 times in PBS/0.05% Tween-20 for 5 mins. Rinse blot in PBS.
12. Develop Blot: For alkaline phosphatase we recommend BCIP/NBT. For horseradish peroxidase we recommend ECL+.
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