Product Specification

                             GAD-Ab ELISA Test Kit

Drug Names

Generic Name: GAD-Ab ELISA Test Kit

INTENDED USE

Detection Kit for Glutamic Acid Decarboxylase Antibody(GAD-Ab)(Enzyme-Linked ImmunoSorbent Assay , ELISA)GAD-Ab EIA kit is used in qualitative tests for Glutamic Acid Decarboxylase Antibody(GAD-Ab)in human serum

SUMMARY AND CLINICAL SIGNIFICANCE

Glutamate decarboxylase (GAD) is a rate-limiting enzyme that converts glutamate into an inhibitory neurotransmitter gamma-aminobutyric acid (GABA), GAD exists in islet β cells, and is also synthesized and secreted by GABA. Glutamate decarboxylase antibody (GADA) is an immune marker for the early stage of type 1 diabetes mellitus and is also used as a therapeutic indicator for the treatment of type 1 diabetes mellitus. The prevalence of GADA in patients with type 1 diabetes mellitus with Graves disease was significantly higher than that in patients with type 1 diabetes without Graves disease. GAD levels in patients with Graves were significantly higher, and the incidence of GADA in non-diabetic patients did not always predict type 1 diabetes.

Product details

PRINCIPLE

GAD-Ab EIA Kit will advance the recombinant human Glutamic Acid Decarboxylase was coated on the reaction plate, is used to capture the GAD-Ab in the sample, washing out extraneous material, then adding horseradish peroxidase (HRP) labeled mouse anti human IgG antibody. Finally, the formation of solid phase labeled anti-human

IgG complex, plus TMB color system after the role of color, the sample can be detected in human serum insulin antibody in the presence of the determination.

TEST PROCEDURE

• Mark the microtitration strips to be used. Set one blank well for background, two wells for negative control, two wells for positive control and other wells for samples.
• Dilute serum samples 1:11 distributing 10ul of serum into 100ul of sample diluent.
• Dispense 100 ul of Negative control as well as Positive control or samples into respective wells.
• Covered the strips with a plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 60 minutes.
• Wash each well for 5 times, 20 seconds each time. (See wash procedure).
• Dispense 100 ul (or two drips) of HRP Conjugate to each well (excluding the blank well) (See drip procedure).
• Covered the strips with a plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 30 minutes.
• Wash each well for 5 times, 10 seconds each time. (See wash procedure).
• Dispense 50 ul (or one drip) of chromogen A to each well (including the blank well).
• Dispense 50 ul (or one drip) of chromogen B to each well (including the blank well).
• Covered the strips with a fresh plate sealer. Mix it gently by swirling the microtiter plate on flat bench. Incubate the plate at 37°C for 15 minutes.
• Dispense 50 ul (or one drip) of stopping solution to each well (including the blank well) and mix completely.
• Read the absorbance of the plate within 10 minutes. (See read procedure)

Important notes

• The kit takes out from the refrigeration environment should be balanced 15-30 minutes in the room temperature then use, ELISA plates coated if has not use up after opened, the plate should be stored in Sealed bag.
• washing buffer will Crystallization separation, it can be heated the water helps dissolve when

dilute . Washing does not affect the result.

• add Sample with sampler Each step, And proofread its accuracy frequently, avoids the experimental error. add sample within 5 min, if the number of sample is much , recommend to use Volley .
• Please make specification curve when you assay, had better make duplicate well, if in the sample the testing material content is excessively high (The sample OD is bigger than the first standard well OD ),please use Sample dilution to dilute certain multiple (n times),then assay. Please multiply total Dilution Times when calculate(×n×5).
• Closure plate membrane only limits the disposable use, in order to avoid the overlapping pollution.
• The substrate please evade the light preservation.
• Please according to use instruction strictly, The test result determination must take the microtiter plate reader as a standard .
• All samples, washing buffer and each kind of reject should according to infective material process.
• This reagent which different batch number component do not mix.
• If it’s different form English instruction, take English instruction as the standard.

Storage and stability

• Store at 2-8℃.

• Seal and return unused reagents to 2-8℃, under which conditions the stability will be retained for 2 months, or until the labeled expiry date, whichever is earlier.