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Product Info Summary
| SKU: | A00713 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-TMEM16A/ANO1 Antibody Picoband®
SKU/Catalog Number
A00713
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-TMEM16A/ANO1 Antibody Picoband® catalog # A00713. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-TMEM16A/ANO1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00713)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human TMEM16A, which shares 83.8% amino acid (aa) sequence identity with both mouse and rat TMEM16A.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00713 is reactive to ANO1 in Human
Observed Molecular Weight
130 kDa
Calculated molecular weight
114.1 kDa
Background of ANO1
Anoctamin-1 (ANO1), also known as oral cancer overexpressed 2 (ORAOV2) or tumor-amplified and overexpressed sequence 2 (TMEM16A), is a protein that in humans is encoded by the ANO1 gene. This gene belongs to a family of membrane proteins containing 8 transmembrane segments, and it is mapped to 11q13.3. ANO1 is a candidate calcium-activated chloride channel that mediates receptor-activated chloride currents in diverse physiologic processes, and it is thought to be responsible for a voltage-sensitive calcium-activated chloride current. Its overexpression was reported in esophageal squamous cell carcinoma and breast cancer progression Crofelemer, an antidiarrhoeal, inhibits this channel. ANO1 has eight transmembrane domains, its pore is large and non-selective, allowing other negatively charged species to permeate.
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.
Application & Images
Applications
A00713 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee
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, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human A431 whole cell, human Hacat whole cell, human PC-3 whole cell, human U251 whole cell
IHC: human oesophagus squama cancer tissue
FCM: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of TMEM16A using anti-TMEM16A antibody (A00713).
Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A431 whole cell lysates,
Lane 2: human Hacat whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human U251 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-TMEM16A antigen affinity purified polyclonal antibody (A00713) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for TMEM16A at approximately 130 kDa. The expected band size for TMEM16A is at 114 kDa.
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IHC analysis of TMEM16A using anti-TMEM16A antibody (A00713).
TMEM16A was detected in paraffin-embedded section of human oesophagus squama cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-TMEM16A Antibody (A00713) 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 # SA1022) with DAB as the chromogen.
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Flow Cytometry analysis of A431 cells using anti-TMEM16A antibody (A00713).
Overlay histogram showing A431 cells stained with A00713 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-TMEM16A Antibody (A00713,1μg/1x106 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-TMEM16A/ANO1 Antibody Picoband® (A00713)
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