Human HBB ELISA Kit (ab235654)
- Datasheet
- References
- Protocols
Overview
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Product name
Human HBB ELISA Kit
See all Hemoglobin kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Serum 8 3.6% -
Sample type
Serum, Cell culture extracts, Hep Plasma, EDTA Plasma, Cit plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
34.1 pg/ml -
Range
0.125 ng/ml - 8 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 102 % - % Cell culture extracts 90 % - % Hep Plasma 101 % - % EDTA Plasma 107 % - % Cit plasma 98 % - % -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
HBB in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of HBB protein in human serum, plasma and cell extracts.
The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
Hemoglobin Subunit Beta (HBB) is a key protein required for transport of oxygen by red blood cells. Hemoglobin is composed of two alpha and beta subunits, and the prosthetic iron containing heme groups. Mutation of the HBB protein is associated with sickle-cell disease and beta thalassemia.
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Tested applications
Suitable for: Sandwich ELISAmore details -
Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human HBB Capture Antibody 1 x 600µl 10X Human HBB Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BI 1 x 6ml Human HBB Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 50ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Involved in oxygen transport from the lung to the various peripheral tissues. -
Tissue specificity
Red blood cells. -
Involvement in disease
Defects in HBA1/HBA2 may be a cause of Heinz body anemias (HEIBAN) [MIM:140700]. This is a form of non-spherocytic hemolytic anemia of Dacie type 1. After splenectomy, which has little benefit, basophilic inclusions called Heinz bodies are demonstrable in the erythrocytes. Before splenectomy, diffuse or punctate basophilia may be evident. Most of these cases are probably instances of hemoglobinopathy. The hemoglobin demonstrates heat lability. Heinz bodies are observed also with the Ivemark syndrome (asplenia with cardiovascular anomalies) and with glutathione peroxidase deficiency.
Defects in HBA1/HBA2 are the cause of alpha-thalassemia (A-THAL) [MIM:604131]. The thalassemias are the most common monogenic diseases and occur mostly in Mediterranean and Southeast Asian populations. The hallmark of alpha-thalassemia is an imbalance in globin-chain production in the adult HbA molecule. The level of alpha chain production can range from none to very nearly normal levels. Deletion of both copies of each of the two alpha-globin genes causes alpha(0)-thalassemia, also known as homozygous alpha thalassemia. Due to the complete absence of alpha chains, the predominant fetal hemoglobin is a tetramer of gamma-chains (Bart hemoglobin) that has essentially no oxygen carrying capacity. This causes oxygen starvation in the fetal tissues leading to prenatal lethality or early neonatal death. The loss of three alpha genes results in high levels of a tetramer of four beta chains (hemoglobin H), causing a severe and life-threatening anemia known as hemoglobin H disease. Untreated, most patients die in childhood or early adolescence. The loss of two alpha genes results in mild alpha-thalassemia, also known as heterozygous alpha-thalassemia. Affected individuals have small red cells and a mild anemia (microcytosis). If three of the four alpha-globin genes are functional, individuals are completely asymptomatic. Some rare forms of alpha-thalassemia are due to point mutations (non-deletional alpha-thalassemia). The thalassemic phenotype is due to unstable globin alpha chains that are rapidly catabolized prior to formation of the alpha-beta heterotetramers.
Note=Alpha(0)-thalassemia is associated with non-immune hydrops fetalis, a generalized edema of the fetus with fluid accumulation in the body cavities due to non-immune causes. Non-immune hydrops fetalis is not a diagnosis in itself but a symptom, a feature of many genetic disorders, and the end-stage of a wide variety of disorders. -
Sequence similarities
Belongs to the globin family. -
Post-translational
modificationsThe initiator Met is not cleaved in variant Thionville and is acetylated. - Information by UniProt
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Alternative names
- 3-prime alpha-globin gene
- A gamma globin
- Alpha 1 globin
see all -
Database links
- Entrez Gene: 3047 Human
- Entrez Gene: 3039 Human
- Entrez Gene: 3040 Human
- Entrez Gene: 3043 Human
- Omim: 141900 Human
- Omim: 141800 Human
- Omim: 142200 Human
- SwissProt: P68871 Human
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Associated products
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Alternative Versions
Applications
Our Abpromise guarantee covers the use of ab235654 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Sandwich ELISA | Use at an assay dependent concentration. |
Images
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Other - Human HBB ELISA Kit (ab235654)SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of human HBB standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native HBB in human serum and plasma samplesThe concentrations of HBB were measured in duplicates, interpolated from the HBB standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 12.5%, plasma (heparin) 5%, plasma (EDTA) 25% and plasma (citrate) 3.13% The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HBB concentration was determined to be 64.9 ng/mL in serum, 28.6 ng/mL in plasma (heparin), 27.1 ng/mL in plasma (EDTA) and 192.2 ng/mL in plasma (citrate).
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Interpolated concentrations of native HBB in TF-1 Extract based on a 37.5 µg/mL extract loadThe concentrations of HBB were measured in duplicate and interpolated from the HBB standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean HBB concentration was determined to be 4.5 ng/mL in TF-1 extract.
Protocols
References
ab235654 has not yet been referenced specifically in any publications.
