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Product Info Summary
| SKU: | A02716-1 |
|---|---|
| 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-Ankyrin erythroid/ANK/ANK1 Antibody Picoband®
SKU/Catalog Number
A02716-1
Size
100 μg/vial
Form
Lyophilized
Description
ANK1 (ankyrin-1) is a core erythrocyte membrane–cytoskeleton adaptor within the ankyrin-1 multiprotein complex that helps maintain red blood cell membrane stability and shape; ANK1 variants are a well-established cause of hereditary spherocytosis (hemolytic anemia). Assay context: Human/Mouse/Rat-reactive antibody tested across ELISA, flow cytometry, IF/ICC, IHC, and WB, enabling localization/abundance mapping of membrane-skeleton phenotypes (e.g., erythroid tissues; interpretation depends on sample context).
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-Ankyrin erythroid/ANK/ANK1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02716-1)
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 Ankyrin erythroid/ANK/ANK1 recombinant protein (Position: N1300-Q1844).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A02716-1 is reactive to ANK1 in Human, Mouse, Rat
Observed Molecular Weight
250 kDa
Calculated molecular weight
206.3 kDa
Background of ANK1
Ankyrin 1, erythrocytic, also known as ANK1, is a protein that in humans is encoded by the ANK1 gene. Ankyrins are a family of proteins that link the integral membrane proteins to the underlying spectrin-actin cytoskeleton and play key roles in activities such as cell motility, activation, proliferation, contact and the maintenance of specialized membrane domains. Multiple isoforms of ankyrin with different affinities for various target proteins are expressed in a tissue-specific, developmentally regulated manner. Most ankyrins are typically composed of three structural domains: an amino-terminal domain containing multiple ankyrin repeats; a central region with a highly conserved spectrin binding domain; and a carboxy-terminal regulatory domain which is the least conserved and subject to variation. Ankyrin 1, the prototype of this family, was first discovered in the erythrocytes, but since has also been found in brain and muscles. Mutations in erythrocytic ankyrin 1 have been associated in approximately half of all patients with hereditary spherocytosis. Complex patterns of alternative splicing in the regulatory domain, giving rise to different isoforms of ankyrin 1 have been described. Truncated muscle-specific isoforms of ankyrin 1 resulting from usage of an alternate promoter have also 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.
Application & Images
Applications
A02716-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, 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, 0.1-0.25 μg/ml, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human HEL whole cell, human K562 whole cell
IHC: human colon cancer tissue, mouse cerebellum tissue, rat cerebellum tissue
ICC/IF: A549 cell
FCM: HEL cell
Validation Images & Assay Conditions
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Western blot analysis of Ankyrin Erythroid/ANK/ANK1 using anti-Ankyrin Erythroid/ANK/ANK1 antibody (A02716-1).
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 HEL whole cell lysates,
Lane 2: human K562 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-Ankyrin Erythroid/ANK/ANK1 antigen affinity purified polyclonal antibody (Catalog # A02716-1) at 0.25 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 Ankyrin Erythroid/ANK/ANK1 at approximately 250 kDa. The expected band size for Ankyrin Erythroid/ANK/ANK1 is at 206 kDa.
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IHC analysis of Ankyrin Erythroid/ANK/ANK1 using anti-Ankyrin Erythroid/ANK/ANK1 antibody (A02716-1).
Ankyrin Erythroid/ANK/ANK1 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 2 μg/ml rabbit anti-Ankyrin Erythroid/ANK/ANK1 Antibody (A02716-1) 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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IHC analysis of Ankyrin Erythroid/ANK/ANK1 using anti-Ankyrin Erythroid/ANK/ANK1 antibody (A02716-1).
Ankyrin Erythroid/ANK/ANK1 was detected in a paraffin-embedded section of mouse cerebellum 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-Ankyrin Erythroid/ANK/ANK1 Antibody (A02716-1) 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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IHC analysis of Ankyrin Erythroid/ANK/ANK1 using anti-Ankyrin Erythroid/ANK/ANK1 antibody (A02716-1).
Ankyrin Erythroid/ANK/ANK1 was detected in a paraffin-embedded section of rat cerebellum 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-Ankyrin Erythroid/ANK/ANK1 Antibody (A02716-1) 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 HEL cells using anti-Ankyrin Erythroid/ANK/ANK1 antibody (A02716-1).
Overlay histogram showing HEL cells stained with A02716-1 (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-Ankyrin Erythroid/ANK/ANK1 Antibody (A02716-1, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IF analysis of Ankyrin Erythroid/ANK/ANK1 using anti-Ankyrin Erythroid/ANK/ANK1 antibody (A02716-1).
Ankyrin Erythroid/ANK/ANK1 was detected in an immunocytochemical section of A549 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-Ankyrin Erythroid/ANK/ANK1 Antibody (A02716-1) 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.
Specific Publications For Anti-Ankyrin erythroid/ANK/ANK1 Antibody Picoband® (A02716-1)
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