The Human Semaphorin 4D (Sema4D) ELISA Kit is specifically designed for the accurate quantification of Semaphorin 4D levels in human serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for various research applications. Semaphorin 4D is a critical protein involved in immune response regulation and cell migration. It plays a significant role in conditions such as inflammation, autoimmune diseases, and cancer, making it a valuable biomarker for studying these diseases and potential therapeutic interventions.
With its user-friendly protocol and reliable performance, the Human Semaphorin 4D (Sema4D) ELISA Kit is an essential tool for researchers looking to investigate the role of Semaphorin 4D in various physiological and pathological processes.
Product Name:
Human Semaphorin 4D/SEMA4D ELISA Kit
SKU:
HUFI01857
Reactivity:
Human
Assay Type:
Sandwich ELISA, Double Antibody
Sensitivity:
75 pg/mL
Range:
125-8000 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)
87-100%
85-105%
92-100%
EDTA Plasma (n = 5)
86-98%
85-99%
82-100%
Heparin Plasma (n = 5)
82-98%
81-90%
85-96%
Recovery:
Sample
Recovery Range (%)
Average (%)
Serum (n = 5)
90-102
96
EDTA Plasma (n = 5)
89-100
97
Heparin Plasma (n = 5)
88-104
92
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.
Biotin-labeled Antibody (Concentrated, 100X)
60 ul
120 ul
2-8°C (Avoid direct light)
HRP-Streptavidin Conjugate (SABC, 100X)
60 ul
120 ul
2-8°C (Avoid direct light)
TMB Substrate
5 ml
10 ml
2-8°C (Avoid direct light)
Sample Dilution Buffer
10 ml
20 ml
2-8°C
Antibody Dilution Buffer
5 ml
10 ml
2-8°C
SABC Dilution Buffer
5 ml
10 ml
2-8°C
Stop Solution
5 ml
10 ml
2-8°C
Wash Buffer(25X)
15 ml
30 ml
2-8°C
Plate Sealer
3 pieces
5 pieces
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Technical Manual
1 copy
1 copy
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Vargovic et al.
Association of Immune Semaphorins with COVID-19 Severity and Outcomes
