The Rat CYP3A4 (Cytochrome P450 3A4) ELISA Kit (SKU: AEFI03638) is a high-sensitivity, 96T sandwich ELISA assay designed for the quantitative detection of CYP3A4 in rat serum, plasma, cell culture supernatant, tissue and cell lysates, and other biological fluids. Utilizing a double antibody sandwich principle, the assay offers a detection range of 78.125–5000 pg/ml with a sensitivity of 46.875 pg/ml and delivers results within 4 hours. The kit includes pre-coated ELISA microplates, lyophilized standards, biotin-labeled antibodies, HRP-streptavidin conjugate (SABC), TMB substrate, and buffers. Assay precision is excellent, with intra- and inter-assay CVs consistently below 6%. The kit is specific to rat CYP3A4, showing no significant cross-reactivity with related proteins, and is traceable to UniProt ID A0A0P6IZE3.
Product Name:
Rat CYP3A4 (Cytochrome P450 3A4) ELISA Kit
SKU:
AEFI03638
Reactivity:
Rat
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
46.875 pg/mL
Range:
78.125-5000 pg/mL
Sample Type:
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
2-8°C for 12 months.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 5)
98-104%
85-103%
86-103%
EDTA Plasma (n = 5)
84-100%
83-94%
85-96%
Heparin Plasma (n = 5)
80-95%
81-99%
88-95%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
85-102
95
EDTA Plasma (n = 5)
85-98
92
Heparin Plasma (n = 5)
88-101
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 reagents and TMB substrate to room temperature. Set standard, test sample and control (zero) wells on the pre-coated plate and record their positions.
2
Primary Incubation: Prepare standards, samples, blanks and load into designated wells. Incubate plate at 37°C for 90 minutes to allow antigen binding.
3
Detection Antibody Binding: Add biotin-labeled detection antibody and incubate at 37°C for 60 minutes.
4
HRP-Streptavidin Binding: Add HRP-Streptavidin (SABC) and incubate at 37°C for 30 minutes.
5
Color Development: Add TMB substrate and incubate in the dark for 10–20 minutes.
6
Stop Reaction & Reading: Add stop solution and measure absorbance at 450 nm immediately.
Sample Type
Protocol
Serum
Allow blood to clot, centrifuge at 1000 × g for 20 minutes, collect supernatant supernatant and store appropriately.
Plasma
Collect using anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C and collect plasma.
Tissue Homogenate
Homogenize tissue in PBS with protease inhibitors, centrifuge and collect supernatant.
Cell Culture Supernatant
Centrifuge at 2500 rpm for 5 minutes and collect clarified supernatant.
Cell Lysate
Lyse cells using lysis buffer with protease inhibitors, centrifuge and collect protein supernatant.
Other Sample Types
For more information about how to process other sample types, (e.g., body fluids, breast milk & more), please contact our Tech Support Team at 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.
