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大鼠（Rat）磷酯酶 Cγ1（PLCγ1） ELISA 检测试剂盒产品说明书
点击次数：422 发布时间：2021/5/10 17:05:56
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Intended use
This PLCγ1 ELISA kit is intended Laboratory for Research use only ａnd is not for
use in diagnostic or
therapeutic procedures.
The S Solution changes the color
from blue to yellow
and the intensity of the
color is measured at 450
nm
using a
spectrophotometer. In order to measure the
concentration of PLCγ1 in
the
sample,
this PLCγ1 ELISA Kit
includes a set
of
calibration standards.
The calibration
standards are assayed at
the same time
as
the
samples ａnd allow
the
operator to
produce a standard curve of Optical Density
versus PLCγ1 concentration. The
activities of PLCγ1 in the samples is then determined by comparing the O.D. of the
samples to the standard curve.
Sample collection ａnd storages
Serum - Use a serum separator tube ａnd allow
samples to clot for 30 minutes
before centrifugation for 10 minutes at approximately 3000×g. Remove serum ａnd
assay immediately or aliquot ａnd store samples at -20℃ or -80℃.Avoid repeated
freeze-thaw cycles
Plasma - Collect plasma
using EDTA or heparin
as an anticoagulant. Centrifuge
samples for 30 minutes at
3000×g at 2-8℃ within
30 minutes of collection. Store
samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.
Cell culture supernates ａnd other biological fluids - Remove particulates
by centrifugation ａnd assay immediately or aliquot ａnd store samples at -20℃or
-80℃. Avoid repeated freeze-thaw cycles.
Note: The samples shoule be centrifugated dequately ａnd no hemolysis or
granule was allowed.
Materials required but not supplied
1.
Standard microplate reader(450nm)
2.
Precision pipettes ａnd Disposable pipette tips.
3.
37 ℃ incubator
Precautions
1.
Do
not substitute reagents from one kit to another. Standard, conjugate ａnd
microplates are matched for optimal performance. Use only the reagents supplied by
manufacturer.
2.
Do not remove microplate from the storage bag until needed. Unused strips
should be stored at 2-8°C in their pouch with the desiccant provided.
3. Mix all reagents before using.
Remove all kit reagents from refrigerator ａnd allow them to reach room temperature( 20-25°C)
Materials supplied
Name
96 determinations
48 determinations
Microelisa stripplate
12*8strips
12*4strips
Standard
0.3ml*6tubes
0.3ml*6tubes
Sample Diluent
6.0ml
3.0ml
HRP-Conjugate reagent
10.0ml
5.0ml
20X Wash solution
25ml
15ml
Chromogen Solution A
6.0ml
3.0ml
Chromogen Solution B
6.0ml
3.0ml
S Solution
6.0ml
3.0ml
Closure plate membrane
2
2
User manual
1
1
Sealed bags
1
1
Note: Standard (S0 → S5)
concentration was
followed by:0,25,50,100,200,400
ng/mL
Reagent preparation
20×wash solution:Dilute with Distilled or deionized water 1:20.
Assay procedure
1.
Prepare all r e a g e n ts
before
starting assay procedure. It is recommended that
all
Standards ａnd Samples be added in duplicate to the Microelisa Stripplate.
2.
Add standard: Set Standard wells, testing sample wells. Add standard 50μl to
standard well.
3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing
sample well; Blank well doesn’t add anyting.
4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip
and incubate for 60 minutes at 37°C.
5. Aspirate each well ａnd wash, repeating the process four times for a total of five
washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle,
manifold
dispenser or autowasher. Complete
removal of liquid
at each step is
essential
to good performance. After the last
wash, remove any
remaining Wash
Solution by aspirating or decanting. Invert the plate ａnd blot it against clean paper
towels.
6.
Add chromogen solution A 50μl ａnd chromogen solution B 50μl to each well.
Gently mix and incubate for 15 minutes at 37°C. Protect from light.
7.
Add 50μl S Solution to each well. The color in the wells should change
from blue to yellow. If the color in the wells is green or the color change does not
appear uniform, gently tap the plate to ensure thorough mixing.
8.
Read the Optical Density (O.D.) at 450
nm using a microtiter plate reader
within 15 minutes.
Calculation of results
1.
This
standard
curve
is used to determine
the amount in an unknown sample.
The
standard
curve
is generated by plotting the average O.D. (450 nm)obtained for each of the six standard concentrations on the vertical
(Y) axis
versus the corresponding concentration on the horizontal (X) axis.
2.
First, calculate the mean O.D. value for each standard
and sample.
All O.D.
values, are subtracted by the mean value of the zero
standard
before result
interpretation.
Construct the
standard curve using graph paper
or statistical
software.
3.
To determine
the amount in
each sample, first locate the O.D.
value on the
Y-axis ａnd extend a horizontal line
to the standard
curve. At
the point of
intersection, draw a vertical line to
the X-axis ａnd
read
the corresponding
concentration.
4.
Any variation in operator, pipetting ａnd washing technique, incubation time or
temperature, ａnd kit age can cause variation in result. Each user should obtain
their own standard curve.
5.
The sensitivity by this assay is 1.0 ng/mL
6.
Standard curve
Storage：
2-8℃.
validity：
six months.
FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR
DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH
ENTIRE PROCEDURE BEFORE BEGINNING!
原创作者：上海隆诚生物科技有限公司
