The 25-Hydroxy Vitamin D3 ELISA Kit from Assay Genie is a highly sensitive and specific assay designed for the quantitative detection of 25-hydroxy vitamin D3 levels in various biological samples, including serum, plasma, and cell culture supernatants. Vitamin D3 is essential for maintaining optimal bone health, immune function, and overall well-being. Deficiency in vitamin D3 has been linked to various health conditions, including osteoporosis, autoimmune diseases, and certain types of cancer.
This ELISA kit offers reliable and reproducible results, making it an ideal tool for researchers and clinicians investigating the role of vitamin D3 in health and disease. With its high sensitivity and accuracy, the 25-Hydroxy Vitamin D3 ELISA Kit from Assay Genie is a valuable resource for studying vitamin D3 metabolism and assessing vitamin D3 status in clinical settings.
Product Name:
25-HVD3 (25-Hydroxy Vitamin D3) ELISA Kit
SKU:
AEFI01010
Reactivity:
Universal
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
0.469 ng/mL
Range:
0.781-50 ng/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)
83-98%
80-98%
80-97%
EDTA Plasma (n = 10)
85-105%
92-105%
86-98%
Heparin Plasma (n = 10)
89-101%
86-103%
86-103%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
85-103
95
EDTA Plasma (n = 10)
87-105
98
Heparin Plasma (n = 10)
94-103
95
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-labeled Antibody (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.
