XbaI ELISA Kit

This kit adopts the double-antibody sandwich ELISA method to detectXbaI content in samples. XbaI-specific antibodies are pre-coated on microplate wells. During detection, standards and samples are added separately into antibody-coated wells.XbaI in samples binds to the immobilized antibodies. After incubation, unbound substances are washed away. HRP-labeledXbaI detection antibody is added to form antibody-antigen-HRP antibody complexes. TMB substrate is added for color development. The color intensity is proportional toXbaI concentration. The residualXbaI in samples is determined by detecting color development ofXbaI standards.Detection Range:0.156~10ng/mlQuantitation Limit:0.156ng/mlLimit of Detection:0.036ng/mlRequired InstrumentsMicroplate reader, microplate constant temperature shaker, vortex mixer, pipettes, plate sealing film, etc.Reagent Preparation1. Equilibration: Allow all required reagents to equilibrate at room temperature (25±3℃) for at least 30 minutes. Determine the number of wells needed, take out required microplate strips, and return unused strips to aluminum foil bags with desiccant. Reseal and store at 2–8℃.2. Prepare 1× Washing Buffer: Dilute 20× Washing Buffer with deionized water at a volume ratio of 1:20 according to required volume, mix well for standby use.3. Standard Solution Dilution (Example):Take 1 vial ofXbaI standard, accurately add 1 mL sample diluent, and vortex to mix thoroughly. The final concentration is 10000 ng/mL, which is used as the standard stock solution.The prepared standard stock solution shall be temporarily stored at 2–8 ℃ and used within 24 hours. For long-term storage, aliquot the solution and keep it at -20 ℃, and use it within 3 months.No.Standard Concentration (ng/ml)Standard Volume (μl)Diluent Volume (μl)Total Volume (μl)Final Concentration (ng/ml)Remaining Volume (μl)StA1000020198020001001950St81005045050010250St7102502505005250St652502505002.5250St52.52502505001.25250St41.252502505000.625250St30.6252502505000.313250St20.3132502505000.156500St1 (Blank)//3003000300Assay Procedure1. Mix all reagents equilibrated to room temperature gently before use.2. Pipette 100 μL of standard solutions (St1~St8) or test samples into corresponding wells. Seal the plate with a plate sealer, then incubate at 25±3 ℃ and 400–500 rpm for 1 hour.3. Washing: Discard the liquid in wells. Add at least 350 μL of 1× Washing Buffer to each well (using a multichannel pipette or wash bottle). Discard residual liquid and pat the plate dry on lint-free absorbent paper. Repeat washing for 3 cycles.(Note: Do not allow wells to dry excessively for a long time before addingXbaI detection antibody. Improper washing may cause high CV between duplicate wells and abnormal absorbance signals.)4. Add 100 μL of HRP-labeledXbaI detection antibody to each well. Seal the plate and incubate at 25±3 ℃, 400–500 rpm for 30 minutes.5. Washing: Discard the liquid in wells. Add at least 350 μL of 1× Washing Buffer to each well. Discard liquid and pat the plate dry. Repeat washing 3 times.(Note: Avoid prolonged excessive drying of wells prior to TMB substrate addition.)6. Color development: Add 100 μL TMB substrate to each well, and incubate for 20 minutes at room temperature in the dark.7. Termination: Add 100 μL Stop Solution to each well, tap the plate gently to mix thoroughly and terminate the reaction.8. Detection: Measure the absorbance at 450 nm within 10 minutes after adding Stop Solution.Data CalculationIt is recommended to perform logarithmic transformation of concentration and fit standard curve with 4-Parameter Logistic regression model.Table 1 Typical Standard Curve DataStandard Concentration (ng/ml)OD Reading 1OD Reading 2Average OD Value102.0822.1022.09251.4741.5131.4932.50.9820.9650.9741.250.6190.6280.6240.6250.4720.4620.4670.3130.3410.3410.3410.1560.3040.280.29200.2540.2520.253Typical Standard Curve (4PL Logistic Model)Performance1. Limit of Detection (LOD): 0.036 ng/ml. Calculated by substituting the average absorbance of 20 blank controls plus 2 standard deviations into the standard curve.2. Limit of Quantitation (LOQ): 0.156 ng/ml. The lowest concentration with coefficient of variation (n=6) ≤ 20%.3. Linearity: Standard curves are plotted with serially diluted standard solutions, with R² ≥ 0.98.4. Repeatability: Evaluated by detecting three samples of known concentrations for 20 replicates.5. Intermediate Precision: Evaluated by detecting three samples of known concentrations 3times by different operators on different days.ParameterRepeatability (n=20)LowConcentrationRepeatability (n=20)MidConcentrationRepeatability (n=20)HighConcentrationIntermediate Precision (n=3)LowConcentrationIntermediate Precision (n=3)MidConcentrationIntermediate Precision (n=)HighConcentrationMean (ng/mL)0.572.397.530.542.367.87SD0.0330.0550.3820.0230.0160.111CV (%)5.72.35.14.30.71.46. Recovery: The recovery rates of spikedXba at different levels within the full detection range in biological matrix were verified.Average Recovery(%)(n=3)Recovery Range (%)94.886.8~100.8Precautions1. For research use only, not for clinical diagnosis or treatment.2. Store reagents as labeled. Equilibrate reagents and samples to room temperature before detection.3. Equilibrate pre-coated microplates to room temperature before opening packages. Return unused strips to sealed bags and store at 2–8℃. Seal all leftover reagents properly.4. MixXbaI standard and HRP detection antibody thoroughly before use.5. Pat washed wells dry on lint-free paper. Do not over-dry wells, which will increase CV and blank OD deviation.6. TMB substrate should be colorless and transparent. Pale blue TMB indicates contamination and cannot be used.7. Samples with extreme pH (<5.0 or >8.5), high salt, polysaccharide, urea, organic solvent or detergent may result in low recovery.8. Due to complex biological matrix, use positive controls and duplicate wells to ensure accurate results.Safety Instructions1. Stop Solution is acidic. Operate with caution.2. Wear proper personal protective equipment: lab coat, gloves, mask, goggles, etc.

Specifications & Purity: This kit adopts the double-antibody sandwich ELISA method to detectXbaI content in samples. XbaI-specific antibodies are pre-coated on microplate wells. During detection, standards and samples are added separately into antibody-coated wells.XbaI in samples binds to the immobilized antibodies. After incubation, unbound substances are washed away. HRP-labeledXbaI detection antibody is added to form antibody-antigen-HRP antibody complexes. TMB substrate is added for color development. The color intensity is proportional toXbaI concentration. The residualXbaI in samples is determined by detecting color development ofXbaI standards.Detection Range:0.156~10ng/mlQuantitation Limit:0.156ng/mlLimit of Detection:0.036ng/mlRequired InstrumentsMicroplate reader, microplate constant temperature shaker, vortex mixer, pipettes, plate sealing film, etc.Reagent Preparation1. Equilibration: Allow all required reagents to equilibrate at room temperature (25±3℃) for at least 30 minutes. Determine the number of wells needed, take out required microplate strips, and return unused strips to aluminum foil bags with desiccant. Reseal and store at 2–8℃.2. Prepare 1× Washing Buffer: Dilute 20× Washing Buffer with deionized water at a volume ratio of 1:20 according to required volume, mix well for standby use.3. Standard Solution Dilution (Example):Take 1 vial ofXbaI standard, accurately add 1 mL sample diluent, and vortex to mix thoroughly. The final concentration is 10000 ng/mL, which is used as the standard stock solution.The prepared standard stock solution shall be temporarily stored at 2–8 ℃ and used within 24 hours. For long-term storage, aliquot the solution and keep it at -20 ℃, and use it within 3 months.No.Standard Concentration (ng/ml)Standard Volume (μl)Diluent Volume (μl)Total Volume (μl)Final Concentration (ng/ml)Remaining Volume (μl)StA1000020198020001001950St81005045050010250St7102502505005250St652502505002.5250St52.52502505001.25250St41.252502505000.625250St30.6252502505000.313250St20.3132502505000.156500St1 (Blank)//3003000300Assay Procedure1. Mix all reagents equilibrated to room temperature gently before use.2. Pipette 100 μL of standard solutions (St1~St8) or test samples into corresponding wells. Seal the plate with a plate sealer, then incubate at 25±3 ℃ and 400–500 rpm for 1 hour.3. Washing: Discard the liquid in wells. Add at least 350 μL of 1× Washing Buffer to each well (using a multichannel pipette or wash bottle). Discard residual liquid and pat the plate dry on lint-free absorbent paper. Repeat washing for 3 cycles.(Note: Do not allow wells to dry excessively for a long time before addingXbaI detection antibody. Improper washing may cause high CV between duplicate wells and abnormal absorbance signals.)4. Add 100 μL of HRP-labeledXbaI detection antibody to each well. Seal the plate and incubate at 25±3 ℃, 400–500 rpm for 30 minutes.5. Washing: Discard the liquid in wells. Add at least 350 μL of 1× Washing Buffer to each well. Discard liquid and pat the plate dry. Repeat washing 3 times.(Note: Avoid prolonged excessive drying of wells prior to TMB substrate addition.)6. Color development: Add 100 μL TMB substrate to each well, and incubate for 20 minutes at room temperature in the dark.7. Termination: Add 100 μL Stop Solution to each well, tap the plate gently to mix thoroughly and terminate the reaction.8. Detection: Measure the absorbance at 450 nm within 10 minutes after adding Stop Solution.Data CalculationIt is recommended to perform logarithmic transformation of concentration and fit standard curve with 4-Parameter Logistic regression model.Table 1 Typical Standard Curve DataStandard Concentration (ng/ml)OD Reading 1OD Reading 2Average OD Value102.0822.1022.09251.4741.5131.4932.50.9820.9650.9741.250.6190.6280.6240.6250.4720.4620.4670.3130.3410.3410.3410.1560.3040.280.29200.2540.2520.253Typical Standard Curve (4PL Logistic Model)Performance1. Limit of Detection (LOD): 0.036 ng/ml. Calculated by substituting the average absorbance of 20 blank controls plus 2 standard deviations into the standard curve.2. Limit of Quantitation (LOQ): 0.156 ng/ml. The lowest concentration with coefficient of variation (n=6) ≤ 20%.3. Linearity: Standard curves are plotted with serially diluted standard solutions, with R² ≥ 0.98.4. Repeatability: Evaluated by detecting three samples of known concentrations for 20 replicates.5. Intermediate Precision: Evaluated by detecting three samples of known concentrations 3times by different operators on different days.ParameterRepeatability (n=20)LowConcentrationRepeatability (n=20)MidConcentrationRepeatability (n=20)HighConcentrationIntermediate Precision (n=3)LowConcentrationIntermediate Precision (n=3)MidConcentrationIntermediate Precision (n=)HighConcentrationMean (ng/mL)0.572.397.530.542.367.87SD0.0330.0550.3820.0230.0160.111CV (%)5.72.35.14.30.71.46. Recovery: The recovery rates of spikedXba at different levels within the full detection range in biological matrix were verified.Average Recovery(%)(n=3)Recovery Range (%)94.886.8~100.8Precautions1. For research use only, not for clinical diagnosis or treatment.2. Store reagents as labeled. Equilibrate reagents and samples to room temperature before detection.3. Equilibrate pre-coated microplates to room temperature before opening packages. Return unused strips to sealed bags and store at 2–8℃. Seal all leftover reagents properly.4. MixXbaI standard and HRP detection antibody thoroughly before use.5. Pat washed wells dry on lint-free paper. Do not over-dry wells, which will increase CV and blank OD deviation.6. TMB substrate should be colorless and transparent. Pale blue TMB indicates contamination and cannot be used.7. Samples with extreme pH (<5.0 or >8.5), high salt, polysaccharide, urea, organic solvent or detergent may result in low recovery.8. Due to complex biological matrix, use positive controls and duplicate wells to ensure accurate results.Safety Instructions1. Stop Solution is acidic. Operate with caution.2. Wear proper personal protective equipment: lab coat, gloves, mask, goggles, etc.

Grade: BioReagent;for Enzyme immunoassay(ELISA)

Storage Temperature: Store at 2-8°C;Protected from light

Shipped in: Wet ice

UPC:

41116126

Weight:

7.06 Ounces

HazmatClass:

No

Quantity:

1

Unit of Measurement:

ST

WeightUOM:

LB

MPN:

X1522789-96T

Country of Origin:

China

Product Size:

96T

Harmonized Tariff Code:

2931.90.90