The 25-Hydroxy Vitamin D3 ELISA Kit from Assay Genie is a highly sensitive and specific assay designed for the precise measurement of 25-hydroxy vitamin D3 levels in serum, plasma, and cell culture supernatants. This kit provides reliable and reproducible results, making it suitable for a variety of research applications.25-hydroxy vitamin D3 is a critical form of vitamin D involved in various physiological processes, including calcium absorption, bone health, and immune function. Deficiencies in 25-hydroxy vitamin D3 have been linked to a variety of health conditions, such as osteoporosis, autoimmune diseases, and cardiovascular disorders, making it an important biomarker for research and clinical studies.
With its easy-to-use protocol and fast results, the 25-Hydroxy Vitamin D3 ELISA Kit is an essential tool for researchers studying vitamin D metabolism, as well as clinicians monitoring vitamin D status in patients. Trust Assay Genie for reliable and accurate measurements of 25-hydroxy vitamin D3 levels in your samples.
Product Name:
25-HVD3 (25-Hydroxy Vitamin D3) ELISA Kit
SKU:
AEFI01032
Reactivity:
Universal
Assay Type:
Competitive ELISA, Coated with Antigen
Sensitivity:
1.875 ng/mL
Range:
3.125-200 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)
85-104%
82-97%
81-97%
EDTA Plasma (n = 10)
86-104%
84-94%
81-100%
Heparin Plasma (n = 10)
90-104%
86-101%
81-98%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 10)
95-102
97
EDTA Plasma (n = 10)
95-104
100
Heparin Plasma (n = 10)
88-104
96
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.
