Product Info Summary
SKU: | A32267-2 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ATP5F1A Antibody Picoband™
SKU/Catalog Number
A32267-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ATP5F1A Antibody Picoband™ catalog # A32267-2. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
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 freezing and thawing.
Cite This Product
Anti-ATP5F1A Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A32267-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ATP5F1A recombinant protein (Position: R12-Q520).
*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
A32267-2 is reactive to ATP5F1A in Human, Mouse, Rat
Applications
A32267-2 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
54 kDa
Calculated molecular weight
59.751kDa
Background of ATP5F1A
ATP synthase F1 subunit alpha, mitochondrial is an enzyme that in humans is encoded by the ATP5F1A gene. This gene encodes a subunit of mitochondrial ATP synthase. Mitochondrial ATP synthase catalyzes ATP synthesis, using 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 consists of three main subunits (a, b, c). This gene encodes the alpha subunit of the catalytic core. Alternatively spliced transcript variants encoding the different isoforms have been identified. Pseudogenes of this gene are located on chromosomes 9, 2, and 16.
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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Assay dilution & Images
Reconsitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/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, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10^6 cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
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 30 ug of sample under reducing conditions.
Lane 1: human SK-OV-3 whole cell lysates,
Lane 2: human MCF-7 whole cell lysates,
Lane 3: human HL-60 whole cell lysates,
Lane 4: human RT4 whole cell lysates,
Lane 5: human Daudi whole cell lysates,
Lane 6: human THP-1 whole cell lysates,
Lane 7: human HepG2 whole cell 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-ATP5F1A antigen affinity purified polyclonal antibody (Catalog # A32267-2) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ATP5F1A at approximately 54 kDa. The expected band size for ATP5F1A is at 54 kDa.
Click image to see more details
Figure 2. Western blot analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
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 30 ug of sample under reducing conditions.
Lane 1: rat heart tissue lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat lung tissue lysates,
Lane 4: rat H9C2(2-1) whole cell lysates,
Lane 5: mouse heart tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse lung tissue lysates,
Lane 8: mouse NIH/3T3 whole cell 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-ATP5F1A antigen affinity purified polyclonal antibody (Catalog # A32267-2) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ATP5F1A at approximately 54 kDa. The expected band size for ATP5F1A is at 54 kDa.
Click image to see more details
Figure 3. IHC analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in a paraffin-embedded section of human endometrial adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 5. IHC analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in a paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 6. IHC analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in a paraffin-embedded section of human thyroid cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 7. IF analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in an immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 8. Flow Cytometry analysis of U937 cells using anti-ATP5F1A antibody (A32267-2).
Overlay histogram showing U937 cells stained with A32267-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATP5F1A Antibody (A32267-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Click image to see more details
Figure 9. IF analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in a paraffin-embedded section of human placenta tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 10. IF analysis of ATP5F1A using anti-ATP5F1A antibody (A32267-2).
ATP5F1A was detected in a paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-ATP5F1A Antibody (A32267-2) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Protein Target Info & Infographic
Gene/Protein Information For ATP5F1A (Source: Uniprot.org, NCBI)
Gene Name
ATP5F1A
Full Name
ATP synthase subunit alpha, mitochondrial
Weight
59.751kDa
Superfamily
ATPase alpha/beta chains family
Alternative Names
ATP synthase subunit alpha, mitochondrial ATP5F1A ATP5A, ATP5A1, ATP5AL2, ATPM, COXPD22, HEL-S-123m, MC5DN4, MOM2, OMR, ORM, hATP1 ATP synthase F1 subunit alpha ATP synthase subunit alpha, mitochondrial|ATP synthase alpha chain, mitochondrial|ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1, cardiac muscle|ATP sythase (F1-ATPase) alpha subunit|epididymis secretory sperm binding protein Li 123m|mitochondrial ATP synthetase, oligomycin-resistant
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on ATP5F1A, check out the ATP5F1A Infographic
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In this infographic, you will see the following information for ATP5F1A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-ATP5F1A Antibody Picoband™ (A32267-2)
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