Categories | Human ELISA Kit |
---|---|
Brand Name: | BT Lab |
Model Number: | Cat.No E6627Hu |
Certification: | CE, ISO9001:2005, MSDS |
Place of Origin: | Shanghai, China |
MOQ: | Negotiation |
Price: | Negotiation |
Supply Ability: | Western Union, T/T |
Delivery Time: | 1-3 business days, bulk order within one week |
Packaging Details: | Wrapped with ice pack and styrofoam package |
Storage: | 2-8°C |
Discount: | Available |
Standard Curve Range: | 7ng/L - 1500ng/L |
Sensitivity: | 4.08ng/L |
OEM: | Acceptable |
Organism Species: | human |
Company Info. |
Shanghai Korain Biotech Co., Ltd |
View Contact Details |
Product List |
Human High Sensitive CNKSR1 ELISA Kit Connector Enhancer of Kinase
Suppressor of Ras 1 ELISA Assay Kit
Cat.No E6627Hu
Precautions
Precision
Intra-Assay Precision (Precision within an assay) Three samples of
known concentration were tested on one plate to assess intra-assay
precision.
Inter-Assay Precision (Precision between assays) Three samples of
known concentration were tested in separate assays to assess
inter-assay precision.
CV(%) = SD/mean x 100
Intra-Assay: CV<8%
Inter-Assay: CV<10%
Assay Principle
This Sandwich ELISA Kit is an Enzyme-Linked Immunosorbent Assay
(ELISA). The plate has been pre-coated with Human CNKSR1 antibody.
CNKSR1 present in the sample is added and binds to antibodies
coated on the wells. And then biotinylated Human CNKSR1 Antibody is
added and binds to CNKSR1 in the sample. Then Streptavidin-HRP is
added and binds to the Biotinylated CNKSR1 antibody. After
incubation unbound Streptavidin-HRP is washed away during a washing
step. Substrate solution is then added and color develops in
proportion to the amount of Human CNKSR1. The reaction is
terminated by addition of acidic stop solution and absorbance is
measured at 450 nm.
Intended Use
This sandwich kit is for the accurate quantitative detection of
Human Connector Enhancer of Kinase Suppressor of Ras 1 (also known
as CNKSR1) in serum, plasma, cell culture supernates, cell lysates,
tissue homogenates.
Reagent Provided
Components | Quantity |
Standard Solution (1600ng/L) | 0.5ml x1 |
Pre-coated ELISA Plate | 12 * 8 well strips x1 |
Standard Diluent | 3ml x1 |
Streptavidin-HRP | 6ml x1 |
Stop Solution | 6ml x1 |
Substrate Solution A | 6ml x1 |
Substrate Solution B | 6ml x1 |
Wash Buffer Concentrate (25x) | 20ml x1 |
Biotinylated Human CNKSR1 Antibody | 1ml x1 |
User Instruction | 1 |
Plate Sealer | 2 pics |
Zipper bag | 1 pic |
Specimen Collection
Serum Allow serum to clot for 10-20 minutes at room temperature.
Centrifuge at 2000-3000 RPM for 20 minutes.
Plasma Collect plasma using EDTA or heparin as an anticoagulant.
Centrifuge samples for 15 minutes at 2000-3000 RPM at 2 - 8°C
within 30 minutes of collection.
Urine Collect by sterile tube. Centrifuge at 2000-3000 RPM for
approximately 20 minutes. When collecting pleuroperitoneal fluid
and cerebrospinal fluid, please follow the procedures
above-mentioned.
Cell Culture Supernatant Collect by sterile tubes when examining secrete components.
Centrifuge at 2000-3000 RPM for approximately 20 minutes. Collect
the supernatants carefully. When examining the components within
the cell, use PBS (pH 7.2-7.4) to dilute cell suspension to the
cell concentration of approximately 1 million/ml. Damage cells
through repeated freeze-thaw cycles to let out the inside
components. Centrifuge at 2000-3000 RPM for approximately 20
minutes.
Tissue and other body fluids Rinse tissues in PBS (pH 7.4) to remove excess blood thoroughly
and weigh before homogenization. Mince tissues and homogenize them
in PBS (pH7.4) with a glass homogenizer on ice. Thaw at 2-8°C or
freeze at -20°C. Centrifuge at 2000-3000 RPM for approximately 20
minutes.
*Sample can't be diluted with this kit. Owing to the the material we use to prepare the kit, the sample
matrix interference may falsely depress the specificity and
accuracy of the assay.
Assay Procedure
1. Prepare all reagents, standard solutions and samples as
instructed. Bring all reagents to room temperature before use. The
assay is performed at room temperature.
2. Determine the number of strips required for the assay. Insert
the strips in the frames for use. The unused strips should be
stored at 2-8°C.
3. Add 50μl standard to standard well. Note: Don’t add antibody to standard well because the standard solution
contains biotinylated antibody.
4. Add 40μl sample to sample wells and then add 10μl anti-CNKSR1
antibody to sample wells, then add 50μl streptavidin-HRP to sample
wells and standard wells (Not blank control well). Mix well. Cover
the plate with a sealer. Incubate 60 minutes at 37°C.
5. Remove the sealer and wash the plate 5 times with wash buffer.
Soak wells with at least 0.35 ml wash buffer for 30 seconds to 1
minute for each wash. For automated washing, aspirate all wells and
wash 5 times with wash buffer, overfilling wells with wash buffer.
Blot the plate onto paper towels or other absorbent material.
6. Add 50μl substrate solution A to each well and then add 50μl
substrate solution B to each well. Incubate plate covered with a
new sealer for 10 minutes at 37°C in the dark.
7. Add 50μl Stop Solution to each well, the blue color will change
into yellow immediately.
8. Determine the optical density (OD value) of each well
immediately using a microplate reader set to 450 nm within 10
minuets after adding the stop solution.