The Vitamin B12 ELISA Kit is a highly sensitive and specific assay designed for the precise quantification of vitamin B12 levels in serum, plasma, and cell culture supernatants. This kit provides accurate and reproducible results, making it an essential tool for research in the fields of nutrition, metabolism, and disease. Vitamin B12, also known as cobalamin, is a vital nutrient that plays a crucial role in DNA synthesis, red blood cell formation, and nerve function. Deficiency in vitamin B12 can lead to a range of health issues, including anemia, neurological problems, and cardiovascular disease.
Therefore, accurate measurement of vitamin B12 levels is essential for assessing nutritional status and monitoring treatment efficacy. Whether studying the impact of diet, aging, or certain medical conditions on vitamin B12 levels, this ELISA kit offers researchers a reliable and efficient method for obtaining valuable data. With its high sensitivity and specificity, the Vitamin B12 ELISA Kit is a valuable tool for advancing understanding of vitamin B12 metabolism and its implications for human health.
Product Name:
VB12 (Vitamin B12) ELISA Kit
SKU:
AEFI01035
Reactivity:
Universal
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
25 pg/mL
Range:
50-500 pg/mL
Sample Type:
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
2-8°C(Sealed), Don't cryopreserve.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 10)
86-97%
81-99%
81-94%
EDTA Plasma (n = 10)
89-104%
85-103%
83-92%
Heparin Plasma (n = 10)
89-100%
86-103%
85-101%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
87-104
96
EDTA Plasma (n = 10)
86-103
94
Heparin Plasma (n = 10)
88-104
94
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1
Reagent & Plate Preparation: Equilibrate TMB substrate for 30 minutes at room temperature. Prepare standards, samples (minimum 1:2 dilution), blanks, assign wells, and pre-wash the plate twice.
2
Sample & Biotin-Antibody Binding: Add 50 µL standard or sample followed by 50 µL biotin-labeled antibody to each well. Mix gently and incubate at 37°C for 45 minutes.
3
Washing: Wash the plate 5 times with wash buffer, allowing 1 minute soak time per wash.
4
Color Development: Add TMB substrate and incubate in the dark at 37°C for 10-20 minutes until color develops.
5
Stop Reaction: Add stop solution to terminate the reaction. The color changes from blue to yellow immediately.
6
Reading: Measure absorbance at 450 nm using a microplate reader.
Sample Type
Protocol
Serum
Allow blood to clot, centrifuge at 1000 × g for 20 minutes, collect supernatant and store appropriately.
Plasma
Collect using EDTA anticoagulant, centrifuge at 1000 × g for 15 minutes at 2–8°C and collect plasma.
Cell Culture Supernatant
Centrifuge at 1000 × g for 20 minutes at 4°C and collect clarified supernatant.
Cell Lysate
Lyse cells using recommended lysis buffer with protease inhibitors, centrifuge at 10,000 rpm for 10 minutes, and collect protein supernatant.
Tissue Homogenate
Homogenize tissue in PBS with protease inhibitors, centrifuge at 5000 × g for 5 minutes, and collect supernatant.
Other Sample Types
Centrifuge samples at 1000 × g for 15 minutes at 2–8°C and collect supernatant. For additional guidance, please contact techsupport@assaygenie.com.
Component
Quantity
Storage
48T
96T
ELISA Microplate (Dismountable)
8×6
8×12
Place the test strips into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
Lyophilized Standard
1 vial
2 vial
Place the standards into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
Biotin-detection Reagent (Lyophilized)
1 vial
1 vial
Place the standards into a sealed foil bag with the desiccant. Store for 1 month at 2-8°C; Store for 12 months at -20°C.
