Human MICB ELISA Kit, Lot 21AO-1098 [Cancer Immune Checkpoint Assay Kit]

CAT#: IOK-05-P1009
Product Type: ELISA Kit
Target: MICB
Short Description
The kit is designed for in vitro quantitative measurement of Human MICB in Plasma, Cell Culture Supernatant, Serum.
Description
Human MICB ELISA Kit for cell culture supernatants, plasma, and serum samples.
Applications
ELISA
Application Notes
Recommended Dilution for serum and plasma samples: 2-fold
Target
MICB
Reactivity
Human
Detection Method
Colorimetric
Method Type
Sandwich ELISA
Analytical Method
Quantitative
Sample Type
Plasma, Cell Culture Supernatant, Serum
Specificity
This ELISA kit shows no cross-reactivity with the following cytokines tested: human Angiogenin, BDNF, BLC, ENA-78, IL-1alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
Components
Pre-Coated 96-well Strip Microplate
Wash Buffer
Stop Solution
Assay Diluent(s)
Lyophilized Standard
Biotinylated Detection Antibody
Streptavidin-Conjugated HRP
TMB One-Step Substrate
Material not included
Distilled or deionized water
Precision pipettes to deliver 2 μL to 1 μL volumes
Adjustable 1-25 μL pipettes for reagent preparation
100 μL and 1 liter graduated cylinders
Tubes to prepare standard and sample dilutions
Absorbent paper
Microplate reader capable of measuring absorbance at 450nm
Log-log graph paper or computer and software for ELISA data analysis
Sensitivity
75 pg/mL
Sample Volume
100 μL
Plate
Pre-coated
Reagent Preparation
1.Bring all reagents and samples to room temperature (18 - 25°C) before use.
2.Sample dilution.
3.Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use.
4.Preparation of standard.
5.Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
6.Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µl of 1x Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate.
7.Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 10,000-fold with 1x Assay Diluent B (Item E).
Assay Procedure
1.Bring all reagents and samples to room temperature (18 - 25°C) before use.
2.Add 100 µl of each standard and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4°C with gentle shaking.
3.Discard the solution and wash 4 times with 1x Wash Solution.
4.Add 100 µl of 1x prepared biotinylated antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
5.Discard the solution. Repeat the wash as in step 3.
6.Add 100 µl of prepared Streptavidin solution to each well. Incubate for 45 minutes at room temperature with gentle shaking.
7.Discard the solution. Repeat the wash as in step 3.
8.Add 100 µl of TMB One-Step Substrate Reagent to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
9.Add 50 µl of Stop Solution to each well. Read at 450 nm immediately.
Calculation of Results
Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Storage
-20 °C
Storage Comment
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Expiry Date
6 months
Note
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Restrictions
For Research Use only
Alternative Name
MHC Class I Polypeptide-Related Sequence B
Synonyms
PERB11.2; MHC class I polypeptide-related sequence B; MHC class I-related protein-like; MICB; LOC512825
Background
The Human MICB (MHC class I Chain-related gene B) ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human MICB in serum, plasma, cell culture supernatants and urine.
Gene ID
4277
UniProt
Q29980
Pathways
Human Leukocyte Antigen (HLA) in Adaptive Immune Response
Protocol
1.Prepare all reagents, samples and standards as instructed in the manual.
2.Add 100 μL of standard or sample to each well.
3.Incubate 2.5 h at RT or O/N at 4°C.
4.Add 100 μL of prepared biotin antibody to each well.
5.Incubate 1 h at RT.
6.Add 100 μL of prepared Streptavidin solution to each well.
7.Incubate 45 min at RT.
8.Add 100 μL of TMB One-Step Substrate Reagent to each well.
9.Incubate 30 min at RT.
10.Add 50 μL of Stop Solution to each well.
11.Read at 450 nm immediately.
For Research Use Only | Not For Clinical Use
Online Inquiry