Rat Activin B (ACVB) (Competitive EIA) ELISA Kit

Mfr.No.

CEA170Ra

Sensitivity

5.29pg/mL

Detection Range

12.35-1,000pg/mL

Reactivity

Rat

Assay Duration

Assay Type

Competitive Inhibition

Description

This assay has high sensitivity and excellent specificity for detection of Activin B (ACVB).
No significant cross-reactivity or interference between Activin B (ACVB) and analogues was observed.

Alternative Names

ACV-B; Activin Beta B Beta B Homodimer

Detection Method

Colormetric

Kit Components

Reagents	Quantity	Reagents	Quantity
Pre-coated, ready to use 96-well strip plate	1	Plate sealer for 96 wells	4
Standard	2	Standard Diluent	1×20mL
Detection Reagent A	1	Assay Diluent A	1×12mL
Detection Reagent B	1×120µL	Assay Diluent B	1×12mL
Reagent Diluent	1×300µL	Stop Solution	1×6mL
TMB Substrate	1×9mL	Instruction manual	1
Wash Buffer (30 × concentrate)	1×20mL

Recovery

Matrices listed below were spiked with certain level of recombinant Activin B (ACVB) and the recovery rates were calculated by comparing the measured value to the expected amount of Activin B (ACVB) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	95-102	99
EDTA plasma(n=5)	98-105	102
heparin plasma(n=5)	80-99	86

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Activin B (ACVB) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1:2	1:4	1:8	1:16
serum(n=5)	80-90%	85-104%	98-105%	91-99%
EDTA plasma(n=5)	90-97%	78-93%	93-104%	87-101%
heparin plasma(n=5)	90-104%	90-98%	78-101%	89-102%

Features

Detects small molecule

Species

Rat

Precautions

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Activin B (ACVB) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Activin B (ACVB) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: C<10%
Inter-Assay: C<12%

Notes

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Activin B (ACVB) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Activin B (ACVB) and unlabeled Activin B (ACVB) (Standards or samples) with the pre-coated antibody specific to Activin B (ACVB). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Activin B (ACVB) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Activin B (ACVB) in the sample.

Usage Notes

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

Protocol

Datasheet

Gene ID	25196
UniProt ID	P17491

Applications	ELISA
Research Area	Endocrinology

Storage	4°C for 1 month/ -20°C for long-term(within shelf life)
Shelf Life	one year

Reference	Activins and their related proteins in colon carcinogenesis: insights from early and advanced azoxymethane rat models of colon cancer.

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ELISA Kit for Activin B (ACVB) (Rat)

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