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Goat Brucella IgG antibody (Brucella-Ab-IgG) ELISA Kit


Species Reactivity

Goat (Capra hircus; Caprine)

UniProt

Abbreviation

Brucella-Ab-IgG

Alternative Names

N/A

Range

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Sensitivity

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Sample Type

Serum, Plasma, Other biological fluids

Detection Method

Competitive ELISA

Analysis Method

Qualitative

Assay Duration

1-3h

Sample Volume

1-200 μL

Detection Wavelengt

450 nm

Cat

AE64524GO

Size

96T 48T

Price

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Availability

3-5 working days

Manual

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Test principle:
This assay employs the competitive enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Brucella-Ab-IgG. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP)-conjugated Brucella-Ab-IgG and incubated. The competitive inhibition reaction is launched between with HRP labeled Brucella-Ab-IgG and unlabeled Brucella-Ab-IgG with the antibody. A substrate solution is added to the wells and the color develops in opposite to the amount of Brucella-Ab-IgG in the sample. The color development is stopped and the intensity of the color is measured.
Product Overview:
Immunoglobulin G (IgG) is a monomeric immunoglobulin, built of two heavy chains γ and two light chains. Each IgG has two antigen binding sites. It is the most abundant immunoglobulin and is approximately equally distributed in blood and in tissue liquids, constituting 75% of serum immunoglobulins in humans. IgG molecules are synthesised and secreted by plasma B cells. IgG antibodies are predominately involved in the secondary antibody response, which occurs approximately one month following antigen recognition, thus the presence of specific IgG generally corresponds to maturation of the antibody response. This is the only isotype that can pass through the human placenta, thereby providing protection to the fetus in utero. Along with IgA secreted in the breast milk, residual IgG absorbed through the placenta provides the neonate with humoral immunity before its own immune system develops.
Components:

Reagents

Quantity

Reagents

Quantity

Assay plate (96 Wells)

1

Instruction manual

1

Standard (lyophilized)

2

Sample Diluent

1 x 20 mL

Biotin-Conjugate (concentrate 100 x)

1 x 120 μL

Biotin-Conjugate Diluent

1 x 20 mL

Streptavidin-HRP  (concentrate 100 x)

1 x 120 μL

Streptavidin-HRP Diluent   

1 x 20 mL

Wash Buffer (concentrate 25 x)

1 x 20 mL

Substrate Solution

1 x 12 mL

Stop Solution

1 x 10 mL

Adhesive Films

4

Specificity:
This assay has high sensitivity and excellent specificity for detection of Goat Brucella-Ab-IgG. No significant cross-reactivity or interference between Goat Brucella-Ab-IgG and analogues was observed.
Recovery:
Matrices listed below were spiked with certain level of recombinant Goat Brucella-Ab-IgG and the recovery rates were calculated by comparing the measured value to the expected amount of Goat Brucella-Ab-IgG in samples.

Sample Type

Number

Recovery range (%)

Average(%)

Serum

10

90-101

96

EDTA plasma

10

89-97

93

Heparin plasma

10

91-99

95

Precision:
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in forty separate assays to assess inter-assay precision. CV (%) = SD/meanX100 Intra-Assay: CV<8% Inter-Assay: CV<12%
Linearity:
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Goat Brucella-Ab-IgG and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample Type

1:2

1:4

1:8

1:16

Serum

78-89%

81-99%

92-103%

95-105%

EDTA plasma

91-101%

90-98%

93-101%

91-98%

Heparin plasma

92-103%

93-102%

92-99%

91-101%

Stability:
The stability of ELISA 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. The loss rate was determined by accelerated thermal degradation test. Keep the kit at 37°C for 4 and 7 days, and compare O.D.values of the kit kept at 37°C with that of at recommended temperature. (referring from China Biological Products Standard, which was calculated by the Arrhenius equation. For ELISA kit, 4 days storage at 37°C can be considered as 6 months at 2 - 8°C, which means 7 days at 37°C equaling 12 months at 2 - 8°C).
Sample collection and storage:
Serum: Use a serum separator tube (SST) and allow samples to clot for two hours at room temperature or overnight at 2 - 8°C before centrifugation for 15 minutes at 1000 × g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Plasma: Collect plasma using EDTA, or heparin as an anticoagulant. Centrifuge for 15 minutes at 1000 × g at 2 - 8°C within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles. Other biological fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.
Kits storage instructions:
Store at 2-8°C. Please refer to Instruction Manual.