Vitamin B-12 ELISA Kit (DEIA-H002)

Regulatory status: For research use only, not for use in diagnostic procedures.

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blood, serum
Species Reactivity
Intended Use
The Quantitative Determination of Vitamin B-12 Concentration in Human Serum by a Microplate Enzyme Immunoassay, Colorimetric.
Contents of Kit
1. Vitamin B-12 Calibrators – 1 mL/vial - Icons A-F
2. Vitamin B-12 Enzyme Reagent – 7.0 mL/vial – Icon
3. Vitamin B-12 Biotin Reagent – 7.0 mL/vial - Icon
4. Streptavidin Coated Plate – 96 wells – Icon
5. Wash Solution Concentrate – 20 mL/vial - Icon
6. Substrate Reagent – 12 mL/vial - Icon
7. Stop Solution – 8 mL/vial - Icon
8. Releasing Agent – 12 mL/vial – Icon
9. Stabilizing Agent – 0.5 mL/vial – Icon
10. Neutralizing Buffer – 7 mL/vial – Icon
Store the kit at 4°C upon receipt. For more detailed information, please download the following document on our website.
Performance Characteristics
1. It is important that the time of reaction in each well is held constant to achieve reproducible results.
2. Pipetting of samples should not extend beyond ten (10) minutes to avoid assay drift.
3. Highly lipemic, hemolyzed or grossly contaminated specimen(s) should not be used.
4. If more than one (1) plate is used, it is recommended to repeat the dose response curve.
5. The addition of substrate solution initiates a kinetic reaction, which is terminated by the addition of the stop solution. Therefore, the substrate and stop solution should be added in the same sequence to eliminate any time-deviation during reaction.
6. Plate readers measure vertically. Do not touch the bottom of the wells.
7. Failure to remove adhering solution adequately in the aspiration or decantation wash step(s) may result in poor replication and spurious results.
8. Use components from the same lot. No intermixing of reagents from different batches.
9. Accurate and precise pipetting, as well as following the exact time and temperature requirements prescribed, are essential. Any deviation from Monobind's IFU may yield inaccurate results.
10. All applicable national standards, regulations and laws, including, but not limited to, good laboratory procedures, must be strictly followed to ensure compliance and proper device usage.
11. It is important to calibrate all the equipment e.g. Pipettes, Readers, Washers and/or the automated instruments used with this device, and to perform routine preventative maintenance.
The within and between assay precision of the Vitamin B-12 ELISA Test System were determined by analyses on three different levels of pool control sera. The number, mean values, standard deviation and coefficient of variation for each of these control sera are presented in Table 2 and Table 3.

*As measured in ten experiments in duplicate over a ten day period.
70.13 pg/mL
General Description
Vitamin B-12 is one of the nine water soluble vitamins important for healthy body functioning. The most important roles Vitamin B-12 plays in the human body are in the formation of red blood cells and the formation of the myelin sheath around the nerves. Since the effects are seen in body systems with a large range of function, the symptoms of Vitamin B-12 deficiency can sometimes be very ambiguous. A deficiency may also take from months to years to manifest depending on the cause and severity. 1, 2, 3 Two of the most common causes of Vitamin B-12 deficiency are diet and age. Because most sources of dietary Vitamin B-12 come from animals, vegans who do not efficiently supplement their diet are at risk. The elderly community is also at high risk because of their diet, as well as the less efficient functioning of their digestive system. 1, 3, 4 Intake of Vitamin B-12 starts by ingestion and then digestion by saliva. Once reaching the gut, Vitamin B-12 bound to proteins in food are released by the acids present. The B12 can then bind the Intrinsic factor. Once bound to IF, Vitamin B-12 is stable enough to travel into the intestines where it can be absorbed into your body through of its association with IF. 1, 5, 6, 7 Two very useful tests to distinguish between Vitamin B-12 deficiency and folate deficiency are methylmalonyl CoA (MMA) and homocysteine (hcy). Both deficiencies are represented by similar symptoms; however, even though both show increased levels of homocysteine, only Vitamin B-12 deficiency causes an increase in methylmalonyl CoA. The increase in levels of methylmalonyl CoA and homocysteine is thought to be the root cause of any symptoms that accompany a Vitamin B-12 deficiency. High levels of these two analytes in the blood stream causes increased oxidative stress to cells therefore causing increased apoptosis. In turn, vascular disease results in the form of atherosclerosis, coronary heart disease and/or neurodegeneration.


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Effects of folinic acid and Vitamin B-12 on ethanol-induced developmental toxicity in mouse


Authors: Xu, Yajun; Li, Li; Zhang, Zhaofeng; Li, Yong

The objective of this study was to assess whether combined supplementation of folinic acid (FA) and Vitamin B-12 (VB12) could suppress ethanol-induced developmental toxicity better than FA alone in mouse embryos cultured in vitro. In this study, exposure to 4.0 mg/ml ethanol for 48 h yielded growth retardation and various malformations of the embryos. FA (10(-5), 10(-4) Mol/l) or VB12 (10(-6), 10(-5) mol/l) alone supplementation improved the growth parameters moderately, however combined supplementation of the two vitamins (10(-5) mol/l FA plus 10(-6) mol/l VB12, 10(-5) mol/l FA plus 10(-5) mol/l VB12, 10(-4) mol/l FA plus 10(-6) Mol/l VB12 and 10(-4) mol/l FA plus 10(-5) mol/l VB12) showed better protective effects, including both the growth and development parameters of the embryos, than either vitamin alone at the same dosage. The present investigation indicated that combined supplementation of folic acid and VB12 might be a better choice than folic acid alone in the prevention of ethanol-induced birth defects. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

pH-responsive controlled release characteristics of solutes with different molecular weights diffusing across membranes of Ca-alginate/protamine/silica hybrid capsules


Authors: He, Fan; Mei, Li; Ju, Xiao-Jie; Xie, Rui; Wang, Wei; Liu, Zhuang; Wu, Fang; Chu, Liang-Yin

Ca-alginate/protamine/silica (APSi) hybrid capsule membranes with pH responsive controlled release characteristics are successfully prepared by combining co-extrusion minifludics, adsorption and biosilicification under different pH conditions from 3 to 7. The microstructures of the prepared capsule membranes are characterized by optical photography, CLSM and SEM. The pH responsive permeability at APSi hybrid capsule membranes is controlled by the electrostatic interactions between Ca-alginate networks and protamine molecules. Four kinds of solute molecules with different molecular weights including methylene blue, VB12, 4kDa and 10kDa FITC-dextran molecules are selected as solute molecules to comparatively study the diffusional permeability characteristics of solutes across APSi hybrid capsule membranes. The results show that, for the solutes with suitable molecule sizes such as VB12 and 4 kDa FITC-dextran, the diffusional permeabilities across the capsule membranes at pH 4 are lower than those at pH 5; however, for the solutes with Loo small molecule size such as methylene blue or Loo large molecule size such as 10 kDa FITC-dextran, the diffusional permeabilities across the capsule membranes at pH 4 are very close to those at pH 5. The results in this study provide valuable guidance for fabrication and application of APSi capsule membranes in the various fields including controlled release of drugs and immobilization of enzymes and so on. (C) 2014 Elsevier BY. All rights reserved.

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