The Human E3 Ubiquitin Protein Ligase ELISA Kit is specifically designed for the quantitative detection of E3 ubiquitin protein ligase levels in human samples, including serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures accurate and reproducible results, making it a valuable tool for a wide range of research applications. E3 ubiquitin protein ligases play a critical role in targeting specific proteins for degradation by the ubiquitin-proteasome system, regulating various cellular processes such as cell cycle progression, DNA repair, and signal transduction. Dysregulation of E3 ligases has been linked to various diseases, including cancer, neurodegenerative disorders, and inflammatory conditions, highlighting the importance of studying their levels and activity.
By using the Human E3 Ubiquitin Protein Ligase ELISA Kit, researchers can gain valuable insights into the role of E3 ligases in disease development and progression, potentially leading to the identification of new therapeutic targets and strategies. Whether studying basic cellular processes or investigating disease mechanisms, this kit provides a reliable and efficient method for measuring E3 ligase levels in human samples.
Product Name:
Human E3 ubiquitin-protein ligase ELISA Kit
SKU:
HUFI01875
Reactivity:
Human
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
0.094 ng/mL
Range:
0.156-10 ng/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)
89-104%
88-98%
89-98%
EDTA Plasma (n = 5)
88-94%
83-100%
80-90%
Heparin Plasma (n = 5)
87-103%
84-97%
91-99%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
91-100
95
EDTA Plasma (n = 5)
87-97
92
Heparin Plasma (n = 5)
87-104
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 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.
