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Rat BAFF/CD257 (B-Cell Activating Factor) ELISA Kit (RTES01025)

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SKU:
RTES01025
$738

Description

Rat BAFF/CD257 (B-Cell Activating Factor) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat BAFF/CD257 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat BAFF/CD257 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat BAFF/CD257, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat BAFF/CD257. The concentration of Rat BAFF/CD257 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityRat
Detection MethodColormetric
Detection Range62.50-4000 pg/mL
Sensitivity37.50 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Rat BAFF/CD257 in samples. No significant cross-reactivity or interference between Rat BAFF/CD257 and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(pg/mL)
O.D Average Corrected
40002.231
2.283
2.2572.177
20001.581
1.593
1.5871.507
10000.943
0.909
0.9260.846
5000.499
0.505
0.5020.422
2500.253
0.229
0.2410.161
1250.184
0.166
0.1750.095
62.500.129
0.129
0.1290.049
00.078
0.082
0.08--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat BAFF/CD257 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat BAFF/CD257 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)198.99492.931413.37179.55507.411400.90
Standard deviation12.0824.5043.399.0128.0175.09
C V (%)6.074.973.075.025.525.36

Recovery

The recovery of Rat BAFF/CD257 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)95-108101
EDTA plasma (n=5)94-106101
Cell culture media (n=5)91-10597

Linearity

Samples were spiked with high concentrations of Rat BAFF/CD257 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)92-10887-9991-104
Average (%)999498
1:4Range (%)87-10184-9884-97
Average (%)949090
1:8Range (%)88-10485-9990-102
Average (%)959295
1:16Range (%)88-10383-9687-100
Average (%)949092

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells ×12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100×)1 vial, 120 uL
Concentrated HRP Conjugate (100×)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25×)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy
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Additional Information

Reactivity:
Rat
ELISA Type:
Sandwich
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