Human IL-1beta ELISA Kit is a highly sensitive assay for researchers seeking to accurately quantify IL-1β (Interleukin 1 Beta) levels in human samples such as serum, plasma, cell culture supernatant, cell or tissue lysate and other liquid samples. The kit uses a sandwich, double antibody ELISA format with a pre-coated microplate, a biotin-labeled detection antibody and an HRP-Streptavidin conjugate, with absorbance read at 450 nm and sample concentrations determined against a standard curve. It offers high sensitivity and excellent specificity for detection of IL-1β, with no significant cross-reactivity or interference between IL-1β and other targets observed. This dual function kit also includes validated Bradford Reagent to quantify total protein concentration for accurate sample normalization.
Product Name:
Human IL-1beta ELISA Kit
SKU:
HUFI03370
Reactivity:
Human
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
2.344 pg/mL
Range:
3.906-250 pg/mL (1pg=140mIU)
Sample Type:
Serum, Plasma, Cell Culture Supernatant, Cell or Tissue Lysate, Other Liquid Samples
Storage:
Store for 1 month at 2-8°C; store for 12 months at -20°C. Date of expiration is on the ELISA Box label.
Linearity:
Sample
1:2
1:4
1:8
Serum (n = 5)
90-98%
88-101%
93-100%
EDTA Plasma (n = 5)
81-98%
85-96%
91-103%
Heparin Plasma (n = 5)
89-103%
85-102%
87-103%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
88-103
96
EDTA Plasma (n = 5)
85-102
94
Heparin Plasma (n = 5)
87-103
97
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 and 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 100 µl into designated wells. Incubate plate at 37°C for 90 minutes to allow antigen binding.
3
Detection Antibody Binding: Wash the plate, then add 100 µl biotin-labeled detection antibody working solution and incubate at 37°C for 60 minutes.
4
HRP-Streptavidin Binding: Wash the plate, then add 100 µl HRP-Streptavidin (SABC) working solution and incubate at 37°C for 30 minutes.
5
Color Development: Wash the plate, then add 90 µl TMB substrate and incubate at 37°C in the dark for 10 to 20 minutes.
6
Stop Reaction and Reading: Add 50 µl stop solution and measure absorbance at 450 nm immediately.
Sample Type
Protocol
Serum
Allow whole blood to coagulate at room temperature (2 h) or 2-8°C overnight, centrifuge at 1000 × g for 20 minutes, collect supernatant and store appropriately.
Plasma
Collect using anticoagulant tubes (EDTA, citrate or heparin), centrifuge within 30 minutes at 1000 × g for 15 minutes at 2 to 8°C and collect plasma.
Tissue Homogenate
Rinse tissue with pre-cooled PBS, homogenize on ice in PBS with protease inhibitors (e.g. 1 mM PMSF), disrupt by ultrasonic homogenization or freeze-thaw, centrifuge at 5000 × g for 5 minutes and collect supernatant. Avoid buffers containing NP-40, Triton X-100 or DTT.
Cell Culture Supernatant
Centrifuge at 2500 rpm for 5 minutes at 2 to 8°C and collect clarified supernatant.
Cell Lysate
Wash cells with pre-cooled PBS, lyse in 0.5-1 ml lysis buffer with protease inhibitors on ice for 30 to 60 minutes, centrifuge at 10,000 rpm for 10 minutes at 2 to 8°C and collect protein supernatant.
Other Sample Types
For more information about how to process other sample types, (e.g., body fluids, breast milk and more), please contact our Tech Support Team at techsupport@assaygenie.com.
Component
Quantity
Storage
48T
96T
ELISA Microplate (Dismountable)
8×6
8×12
Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 12 months at -20°C.
Lyophilized Standard
1 vial
2 vials
Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 12 months at -20°C.
