추천 제품
형태
liquid
Quality Level
미포함
preservative
제조업체/상표
Calbiochem®
저장 조건
OK to freeze
protect from light
색상
clearvery light blue
배송 상태
ambient
저장 온도
2-8°C
InChI
1S/C16H20N2/c1-9-5-13(6-10(2)15(9)17)14-7-11(3)16(18)12(4)8-14/h5-8H,17-18H2,1-4H3
InChI key
UAIUNKRWKOVEES-UHFFFAOYSA-N
일반 설명
Peroxidase substrate that produces a blue soluble end product that can be read at 370 nm or 650 nm. Use of stop solution enhances sensitivity two- to four-fold and results in a yellow solution that can be read at 450 nm.
Soluble TMB substrate supplied as a single component system.
경고
Toxicity: Irritant (B)
물리적 형태
Single component system containing 1.46 mM TMB, 2.21 mM H₂O₂ in a proprietary solvent, pH 3.1 ± 0.5.
재구성
Protect from exposure to direct sunlight. Discard if the solution turns blue or turbid. The reagent is stable at room temperature, but we recommend storage in the refrigerator for longer shelf life. Warm to assay temperature before use.
기타 정보
Product tested for stability at 4°C and 18-26°C. Product performance is tested on the final product by ELISA.
Suggested Procedure for Use of TMB in HRP-based ELISAs:
1. Complete all required incubations with antibodies and HRP-labeled probes.
2. Wash plate wells at least 4 times with phosphate-buffered saline (PBS) or Tris-buffered saline (TBS), containing 0.1% Tween®-20 detergent.
3. After the final wash, shake and blot all residual buffer from the wells.
4. Add 100 μl TMB solution to each well and incubate for 5-30 min.
5. Options for measurements:
a. For kinetic assays, the reaction can be monitored as a function of time by reading absorbance at 650 nm at intermediate intervals.
b. For endpoint assays that preserve the blue chromogen, the reaction should be stopped by addition of 100 µl of 0.1% sodium fluoride (NaF) and the absorbance read at 650 nm.
c. If increased sensitivity is desired for endpoint assays, the reaction should be stopped by addition of 100 μl of either 500 mM H2SO4 or 250 mM HCl and the absorbance read at 450 nm WITHIN 5 MIN. Addition of acid converts the blue radical to the yellow diimine, which absorbs at 450 nm.
NOTE: Optimal incubation times may vary depending on the amount of HRP present. If color develops too quickly, zero-order kinetics will not prevail. Dilution of the probe, antibody, or HRP-labeled reagent may be required. Variations in time, reagent volumes, and temperature may also require further standardization by the user.
Suggested Procedure for Use of TMB in HRP-based ELISAs:
1. Complete all required incubations with antibodies and HRP-labeled probes.
2. Wash plate wells at least 4 times with phosphate-buffered saline (PBS) or Tris-buffered saline (TBS), containing 0.1% Tween®-20 detergent.
3. After the final wash, shake and blot all residual buffer from the wells.
4. Add 100 μl TMB solution to each well and incubate for 5-30 min.
5. Options for measurements:
a. For kinetic assays, the reaction can be monitored as a function of time by reading absorbance at 650 nm at intermediate intervals.
b. For endpoint assays that preserve the blue chromogen, the reaction should be stopped by addition of 100 µl of 0.1% sodium fluoride (NaF) and the absorbance read at 650 nm.
c. If increased sensitivity is desired for endpoint assays, the reaction should be stopped by addition of 100 μl of either 500 mM H2SO4 or 250 mM HCl and the absorbance read at 450 nm WITHIN 5 MIN. Addition of acid converts the blue radical to the yellow diimine, which absorbs at 450 nm.
NOTE: Optimal incubation times may vary depending on the amount of HRP present. If color develops too quickly, zero-order kinetics will not prevail. Dilution of the probe, antibody, or HRP-labeled reagent may be required. Variations in time, reagent volumes, and temperature may also require further standardization by the user.
법적 정보
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
TWEEN is a registered trademark of Croda International PLC
신호어
Warning
유해 및 위험 성명서
Hazard Classifications
Acute Tox. 4 Oral - Aquatic Chronic 4 - Carc. 2
Storage Class Code
11 - Combustible Solids
WGK
WGK 3
Flash Point (°F)
Not applicable
Flash Point (°C)
Not applicable
시험 성적서(COA)
제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.
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