Transforming growth factor beta (TGF-β) is a multifunctional cytokine that plays a critical role in various biological processes, including cell growth, differentiation, and immune regulation. It is part of a superfamily of proteins known for their ability to control cellular functions and influence tissue development. TGF-β is involved in numerous physiological and pathological processes, such as embryogenesis, wound healing, inflammation, and cancer progression. By measuring the levels of TGF-β in biological samples, such as serum or cell culture supernatants, the TGF Beta ELISA kit provides a valuable tool for researchers and clinicians to study the role of TGF-β in disease mechanisms, monitor treatment responses, and explore potential therapeutic interventions.
This immunoassay kit allows for the in vitro quantitative determination of Human TGF-beta1 concentrations in serum plasma and other biological fluids.
Sensitivity:
18.75 pg/mL
Range:
31.25-2000 pg/mL (1pg=25mIU)
Storage:
4°C for 6 months
Note:
For Research Use Only
Recovery:
Matrices listed below were spiked with certain level of Human TGF-beta1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human TGF-beta1 in samples.
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
85-102
95
EDTA Plasma (n = 5)
96-104
99
UFH Plasma (n = 5)
92-102
97
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human TGF-beta1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample
1:2
1:4
1:8
Serum (n = 5)
89-95%
89-103%
85-103%
EDTA Plasma (n = 5)
92-101%
86-101%
86-95%
UFH Plasma (n = 5)
81-97%
80-99%
85-100%
CV(%):
Intra-Assay: CV<8% Inter-Assay: CV<10%
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. Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
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. Wash plate 2 times before adding standard, sample and control (zero) wells.
2
Primary Incubation: Aliquot 0.1 ml standard solutions and samples into designated wells. Incubate plate at 37°C to allow antigen binding.
3
Detection Antibody Binding: Add biotin-labeled detection antibody and incubate at 37°C.
4
HRP-Streptavidin Binding: Add HRP-Streptavidin (SABC) and incubate at 37°C.
5
Color Development: Add TMB substrate and incubate in the dark.
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 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
ELISA Microplate (Dismountable)
8×12 strips
4°C for 6 months
Lyophilized Standard
2 vials
4°C / -20°C
Sample/Standard Dilution Buffer
20 ml
4°C
Biotin-labeled Antibody (Concentrated)
120 ul
4°C (Avoid direct light)
Antibody Dilution Buffer
10 ml
4°C
HRP-Streptavidin Conjugate (SABC)
120 ul
4°C (Avoid direct light)
SABC Dilution Buffer
10 ml
4°C
TMB Substrate
10 ml
4°C (Avoid direct light)
Stop Solution
10 ml
4°C
Wash Buffer (25X)
30 ml
4°C
Plate Sealer
5 pieces
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Other materials required:
Microplate reader with 450 nm wavelength filter
Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
