人 SULT1B1 ELISA Pair Set

ABIN2010513 产品详细信息, 供应商: Log in to see
蛋白/底物
  • ST1B1
  • ST1B2
  • SULT1B2
  • St2b2
  • SULT1B
  • cSULT1B1
  • MGC80677
  • sulfotransferase family, cytosolic, 1B, member 1
  • sulfotransferase family 1B, member 1
  • sulfotransferase family, cytosolic, 1D, member 1
  • SULT1B1
  • Sult1b1
  • sult1b1
  • SULT1D1
适用

应用范围
ELISA
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原理 The ELISA Pair Set is a solid phase sandwich ELISA (Enzyme-Linked Immunosorbent Assay). It utlizes monoclonal antibody specific for SULT1B1 coated on a 96-well plate. Standards and samples are added to the wells,and any SULT1B1 present binds to the immobilized antibody. The wells are washed and a horseradish peroxidaseconjugated mouse anti-SULT1B1 monoclonal antibody is then added, producing an antibody-antigen-antibody"sandwich". The wells are again washed and TMB substrate solution is loaded, which produces color in proportion tothe amount of SULT1B1 present in the sample. To end the enzyme reaction, the stop solution is added andabsorbances of the microwell are read at 450 nm.
灵敏度 The minimum detectable dose of Human SULT1B1 was determined to be approximately 156.25 pg/mL. This is defined as at least three times standard deviations above the mean optical density of 10 replicates of the zero standard.
产品特性 Pair Set
组件 Capture Antibody - 0.3 mg/mL of mouse anti-SULT1B1 monoclonal antibody. Dilute to a workingconcentration of 2.0 μg/mL in CBS before coating.
Detection Antibody - 0.5 mg/mL mouse anti-SULT1B1 monoclonal antibody conjugated tohorseradish-peroxidase (HRP) . Dilute to working concentration of 0.5 μg/mL in detection antibodydiluteion buffer before use.
Standard - Each vial contains 390 ng of recombinant SULT1B1. Reconstitute standard powderwith 1mL detection antibody dilution buffer. After reconstitution, store at -20 °C to -70 °C in a manualdefrost freezer. A seven-point standard curve using 2-fold serial dilutions in sample dilution buffer,and a high standard of 10000 pg/mL is recommended.
试剂未包括 CBS - 0.05 M
Na2CO3 - 0.05 M
NaHCO3, pH 9.6, 0.2 μm filtered
TBS - 25 mM
Tris, adjust pH to 7.4 by HCl
Wash Buffer - 0.05 % Tween20 in TBS, pH 7.2 - 7.4
Blocking Buffer - 5 % BSA in Wash Buffer
Sample dilution buffer - 0.5 % BSA in wash buffer, pH 7.2 - 7.4, 0.2 μm filtered
Detection antibody dilution buffer - 0.5 % BSA in wash buffer, pH 7.2 - 7.4, 0.2 μm filtered
Substrate Solution: To achieve best assay results, fresh substrate solution is recommended Substrate stock solution - 10 mg/mL TMB (Tetramethylbenzidine ) in DMSO Substrate dilution buffer - 0.05 M Na2HPO4 and 0.025 M citric acid , adjust pH to 5.5
Substrate working solution - For each plate dilute 250 μL substrate stock solution in 25 mL substrate dilutionbuffer and then add 80 μL 0.75 % H2O2 , mix it well
Stop Solution - 2 N H2SO4
别名 SULT1B1
背景 Sulfotransferase family cytosolic 1B member 1, also known as Sulfotransferase 1B1, Sulfotransferase 1B2, Thyroidhormone sulfotransferase, SULT1B1 and ST1B2, is a cytoplasm protein which belongs to thesulfotransferase 1 family. Sulfotransferase enzymes catalyze the sulfate conjugation of many hormones, neurotransmitters, drugs, and xenobioticcompounds. These cytosolic enzymes are different in their tissue distributions and substrate specificities. SULT1B1 ishighly expressed in the liver, peripheral blood leukocytes, colon (mucosal lining) , small intestine (jejunum) and spleen. A lesser expression of SULT1B1 was observed in the lung, placenta and thymus. SULT1B1 catalyzes the sulfateconjugation of many hormones, neurotransmitters, drugs and xenobiotic compounds. Sulfonation increases the watersolubility of most compounds, and therefore their renal excretion, but it can also result in bioactivation to form activemetabolites. SULT1B1 sulfates dopamine, small phenols such as 1-naphthol and p-nitrophenol and thyroid hormones, including 3, 3'-diiodothyronine, triidothyronine, reverse triiodothyronine and thyroxine.
研究领域 Signaling, Metabolism, Hormones
应用备注 Optimal working dilution should be determined by the investigator.
说明

The Human SULT1B1 ELISA Pair Set is for the quantitative determination of Human SULT1B1.This ELISA Pair Set contains the basic components required for the development of sandwich ELISAs.

试剂准备
  1. Dilute the capture antibody to the working concentration in CBS. Immediately coat a 96-well microplate with 100 μL per well of the diluted capture antibody. Seal the plate and incubate overnight at 4 °C.
    2. Aspirate each well and wash with at least 300 μL wash buffer, repeating the process two times for a total of three washes. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining wash buffer by inverting the plate and blotting it against clean paper towels.
    3. Block plates by adding 300 μL of blocking buffer to each well. Incubate at room temperature for a minimum of 1 hour.
    4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
实验流程
  1. Add 100 μL of sample or standards in sample dilution buffer per well. Seal the plate and incubate 2 hours at roomtemperature.
    2. Repeat the aspiration/wash as in step 2 of plate preparation.
    3. Add 100 μL of the detection antibody, diluted in antibody dilution buffer, to each well. Seal the plate and incubate 1hour at room temperature.
    4. Repeat the aspiration/wash as in step 2 of plate preparation.
    5. Add 200 μL of substrate solution to each well. Incubate for 20 minutes at room temperature (if substrate solutionis not as requested, the incubation time should be optimized). Avoid placing the plate in direct light.
    6. Add 50 μL of stop solution to each well. Gently tap the plate to ensure thorough mixing.
    7. Determine the optical density of each well immediately, using a microplate reader set to 450 nm.
结果分析

Calculate the mean absorbance for each set of duplicate standards, controls and samples. Subtract the mean zero standard absorbance from each. Construct a standard curve by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and draw a best fit curve through the points on the graph. To determine the concentration of the unknowns, find the unknowns' mean absorbance value on the y-axis and draw a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the x-axis and read the concentration. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor. Alternatively, computer-based curve-fitting statistical software may also be employed to calculate the concentration of the sample.

限制 仅限研究用
状态 Lyophilized
注意事项 Avoid repeated freeze-thaw cycles.
储存条件 -20 °C/-80 °C
储存方法 Capture Antibody: Aliquot and store at -20°C to -80°C for up to 6 months from date of receipt. Detection Antibody: Protect it from prolonged exposure to light. Aliquot and store at -20°C to -80°C and for up to 6 months from date of receipt. Standard: Store lyophilized standard at -20°C to -80°C for up to 6 months from date of receipt. Aliquot and store the reconstituted Standard at -80°C for up to 1 month.
有效期 6 months
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