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ab290021 is the same product as ab110322.

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Images

Immunocytochemistry - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (AB290021), expandable thumbnail
  • Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (AB290021), expandable thumbnail
  • Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (AB290021), expandable thumbnail

Key facts

Reactive species
Human, Caenorhabditis elegans, Mouse, Cow, Rat

Reactivity data

Application
WB
Reactivity
Reacts
Dilution info
1 µg/mL
Notes

-

Application
ICC
Reactivity
Reacts
Dilution info
10 µg/mL
Notes

-

Target data

Function

ADP:ATP antiporter that mediates import of ADP into the mitochondrial matrix for ATP synthesis, and export of ATP out to fuel the cell (PubMed:21586654, PubMed:27693233). Cycles between the cytoplasmic-open state (c-state) and the matrix-open state (m-state): operates by the alternating access mechanism with a single substrate-binding site intermittently exposed to either the cytosolic (c-state) or matrix (m-state) side of the inner mitochondrial membrane (By similarity). In addition to its ADP:ATP antiporter activity, also involved in mitochondrial uncoupling and mitochondrial permeability transition pore (mPTP) activity (PubMed:31883789). Plays a role in mitochondrial uncoupling by acting as a proton transporter: proton transport uncouples the proton flows via the electron transport chain and ATP synthase to reduce the efficiency of ATP production and cause mitochondrial thermogenesis (By similarity). Proton transporter activity is inhibited by ADP:ATP antiporter activity, suggesting that SLC25A4/ANT1 acts as a master regulator of mitochondrial energy output by maintaining a delicate balance between ATP production (ADP:ATP antiporter activity) and thermogenesis (proton transporter activity) (By similarity). Proton transporter activity requires free fatty acids as cofactor, but does not transport it (By similarity). Also plays a key role in mPTP opening, a non-specific pore that enables free passage of the mitochondrial membranes to solutes of up to 1.5 kDa, and which contributes to cell death (PubMed:31883789). It is however unclear if SLC25A4/ANT1 constitutes a pore-forming component of mPTP or regulates it (By similarity). Acts as a regulator of mitophagy independently of ADP:ATP antiporter activity: promotes mitophagy via interaction with TIMM44, leading to inhibit the presequence translocase TIMM23, thereby promoting stabilization of PINK1 (By similarity).

Additional Targets

SLC25A5

Alternative names

AAC1, ANT1, SLC25A4, ADP/ATP translocase 1, Adenine nucleotide translocator 1, Solute carrier family 25 member 4, ANT 1

What's included?

100 µg
Components
Anti-Adenine Nucleotide Translocase (ANT) monoclonal antibody
1 x 100 µg
Incubation Solution
1 x 100 mL

Recommended products

ab290021 is the same product as ab110322.

Key facts

Reactive species
Human, Caenorhabditis elegans, Mouse, Cow, Rat

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

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 notbe 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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3 product images

  • Immunocytochemistry - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (ab290021), expandable thumbnail

    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.

  • Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (ab290021), expandable thumbnail

    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 (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] (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.

  • Western blot - Anti-Adenine Nucleotide Translocator 1 + 2/ANT 1 + 2 antibody [5F51BB5AG7] (ab290021), expandable thumbnail

    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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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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