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Product Info Summary
| SKU: | A00233-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-Hemoglobin/HBA1/HBA2 Antibody Picoband®
SKU/Catalog Number
A00233-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hemoglobin/HBA1/HBA2 Antibody Picoband® catalog # A00233-1. 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-Hemoglobin/HBA1/HBA2 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00233-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Hemoglobin recombinant protein (Position: V2-R142). Human Hemoglobin shares 85.8% amino acid (aa) sequence identity with mouse Hemoglobin.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
A00233-1 is reactive to HBA1 in Human
Observed Molecular Weight
15 kDa
Calculated molecular weight
15.3 kDa
Background of HBA1
The human alpha globin gene cluster located on chromosome 16 spans about 30 kb and includes seven loci: 5'- zeta - pseudozeta - mu - pseudoalpha-1 - alpha-2 - alpha-1 - theta - 3'. The alpha-2 (HBA2) and alpha-1 (HBA1) coding sequences are identical. These genes differ slightly over the 5' untranslated regions and the introns, but they differ significantly over the 3' untranslated regions. Two alpha chains plus two beta chains constitute HbA, which in normal adult life comprises about 97% of the total hemoglobin; alpha chains combine with delta chains to constitute HbA-2, which with HbF (fetal hemoglobin) makes up the remaining 3% of adult hemoglobin. Alpha thalassemias result from deletions of each of the alpha genes as well as deletions of both HBA2 and HBA1; some nondeletion alpha thalassemias have also been reported.
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
A00233-1 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), 2-5μg/ml, Human
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human placenta tissue
IHC: human placenta tissue, human tonsil tissue
FCM: K562 cell
Validation Images & Assay Conditions
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Western blot analysis of Hemoglobin using anti-Hemoglobin antibody (A00233-1).
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 placenta tissue 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-Hemoglobin antigen affinity purified polyclonal antibody (Catalog # A00233-1) 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 Hemoglobin at approximately 15 kDa. The expected band size for Hemoglobin is at 15 kDa.
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IHC analysis of Hemoglobin using anti-Hemoglobin antibody (A00233-1).
Hemoglobin 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-Hemoglobin Antibody (A00233-1) 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.
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IHC analysis of Hemoglobin using anti-Hemoglobin antibody (A00233-1).
Hemoglobin was detected in a paraffin-embedded section of human tonsil 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-Hemoglobin Antibody (A00233-1) 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.
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Flow Cytometry analysis of K562 cells using anti-Hemoglobin antibody (A00233-1).
Overlay histogram showing K562 cells stained with A00233-1 (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-Hemoglobin Antibody (A00233-1, 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 (Red line) was also used as a control.
Specific Publications For Anti-Hemoglobin/HBA1/HBA2 Antibody Picoband® (A00233-1)
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1 Customer Q&As for Anti-Hemoglobin/HBA1/HBA2 Antibody Picoband®
Question
We are currently using anti-Hemoglobin/HBA1/HBA2 antibody A00233-1 for rat tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, rat. Is it true that the antibody can work on primate tissues as well?
J. Miller
Verified customer
Asked: 2013-06-17
Answer
The anti-Hemoglobin/HBA1/HBA2 antibody (A00233-1) has not been validated for cross reactivity specifically with primate tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in primate you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-06-17
