Anti-ATP5F1,2,3/ATP5MC1,2,3 Picoband™ Antibody (monoclonal, 12E9)

Product Name

Anti-ATP5F1,2,3/ATP5MC1,2,3 Picoband™ Antibody (monoclonal, 12E9)

View all ATP synthase C mature Antibodies

SKU/Catalog Number

M09735

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-ATP5F1,2,3/ATP5MC1,2,3 Picoband™ Antibody (monoclonal, 12E9) catalog # M09735. Tested in Flow Cytometry, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat.

Storage & Handling

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.

Cite This Product

Anti-ATP5F1,2,3/ATP5MC1,2,3 Picoband™ Antibody (monoclonal, 12E9) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M09735)

Host

Mouse

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Monoclonal

Clone Number

12E9

Isotype

Mouse IgG2b

Immunogen

E.coli-derived human ATP5G1,2,3/ATP5MC1,2,3 recombinant protein (Position: D62-L113).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M09735 is reactive to ATP5MC1 in Human, Monkey, Mouse, Rat

Applications

M09735 is guaranteed for Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

10-14 kDa

Calculated molecular weight

14.277kDa

Background of ATP synthase C mature

The ATP5MC1 gene is one of three human paralogs that encode membrane subunit c of the mitochondrial ATP synthase. It is mapped to 17q21.32. This gene encodes a subunit of mitochondrial ATP synthase. Mitochondrial ATP synthase catalyzes ATP synthesis, utilizing an electrochemical gradient of protons across the inner membrane during oxidative phosphorylation. ATP synthase is composed of two linked multi-subunit complexes: the soluble catalytic core, F1, and the membrane-spanning component, Fo, comprising the proton channel. The catalytic portion of mitochondrial ATP synthase consists of 5 different subunits (alpha, beta, gamma, delta, and epsilon) assembled with a stoichiometry of 3 alpha, 3 beta, and a single representative of the other 3. The proton channel seems to have nine subunits (a, b, c, d, e, f, g, F6 and 8). This gene is one of three genes that encode subunit c of the proton channel. Each of the three genes have distinct mitochondrial import sequences but encode the identical mature protein. Alternatively spliced transcript variants encoding the same protein have been identified.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat, Monkey
Flow Cytometry, 1-3μg/1x10⁶ cells, Human, Mouse, Rat

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of ATP5F1,2,3/ATP5MC1,2,3 using anti-ATP5F1,2,3/ATP5MC1,2,3 antibody (M09735).
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 HL-60 whole cell lysates,
Lane 2: monkey COS-7 whole cell lysates,
Lane 3: rat kidney tissue lysates,
Lane 4: rat heart 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 mouse anti-ATP5F1,2,3/ATP5MC1,2,3 antigen affinity purified monoclonal antibody (Catalog # M09735) at 0.5 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 # EK1001) with Tanon 5200 system. A specific band was detected for ATP5F1,2,3/ATP5MC1,2,3 at approximately 10-14KD. The expected band size for ATP5F1,2,3/ATP5MC1,2,3 is at 10-14KD.

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Figure 2. Flow Cytometry analysis of HEPA1-6 cells using anti-ATP5F1,2,3/ATP5MC1,2,3 antibody (M09735).
Overlay histogram showing HEPA1-6 cells stained with M09735 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ATP5F1,2,3/ATP5MC1,2,3 Antibody (M09735, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 3. Flow Cytometry analysis of HEPG2 cells using anti-ATP5F1,2,3/ATP5MC1,2,3 antibody (M09735).
Overlay histogram showing HEPG2 cells stained with M09735 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ATP5F1,2,3/ATP5MC1,2,3 Antibody (M09735, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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Figure 4. Flow Cytometry analysis of RH35 cells using anti-ATP5F1,2,3/ATP5MC1,2,3 antibody (M09735).
Overlay histogram showing RH35 cells stained with M09735 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ATP5F1,2,3/ATP5MC1,2,3 Antibody (M09735, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For ATP5MC1 (Source: Uniprot.org, NCBI)

Alternative Names

ATP synthase, H+ transporting, mitochondrial F0 complex, subunit c (subunit 9); ATP synthase, H+ transporting, mitochondrial Fo complex, subunit C1 (subunit 9); ATP5A; ATP5G; ATPase protein 9; ATPase subunit 9; H+ transporting, mitochondrial F0 complex, subunit C1 (subunit 9); isoform 1; mitochondrial ATP synthase, subunit 9; mitochondrial ATP synthase, subunit C; mitochondrial ATP5MC1 ATP5A, ATP5G, ATP5G1 ATP synthase membrane subunit c locus 1 ATP synthase F(0) complex subunit C1, mitochondrial|ATP synthase lipid-binding protein, mitochondrial|ATP synthase proteolipid P1|ATP synthase proton-transporting mitochondrial F(0) complex subunit C1|ATP synthase subunit 9|ATP synthase, H+ transporting, mitochondrial F0 complex, subunit C1 (subunit 9)|ATP synthase, H+ transporting, mitochondrial F0 complex, subunit c (subunit 9)|ATP synthase, H+ transporting, mitochondrial Fo complex subunit C1 (subunit 9)|ATPase protein 9|ATPase subunit 9|ATPase subunit C|dicyclohexylcarbodiimide (DCCD)-reactive proteolipid subunit|mitochondrial ATP synthase, subunit 9|mitochondrial ATP synthase, subunit C

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