TMB（3，3'，5，5'-四甲基聯(lián)苯胺）是辣根過(guò)氧化物酶（HRP）的發(fā)色底物。 在HRP對過(guò)氧化氫進(jìn)行酶促降解過(guò)程中，TMB產(chǎn)生深藍色，加入終止液產(chǎn)生清晰的黃色，在450nm處測得。 與OPD（鄰苯二胺二氯化物）相比，該底物可提供更高的信號并在ELISA中具有更高的靈敏度。

分析：對比分析底物的活性，背景，顏色和pH值是否一致。

背景：將100μl底物加到標準微量滴定板的8個(gè)孔中，然后加入100μl 0.2 M硫酸。 在450 nm處讀取背景吸光度。
活性： 將100μl底物添加到每個(gè)孔中，并孵育13分鐘。 加入100 μl 0.2 M硫酸終止反應。 在450nm處讀取吸光度。 將活性與具有已知活性的標準TMB進(jìn)行比較。

產(chǎn)品特性：活性90 - 110%。背景 Abs. < 0.025。顏色：透明溶液。pH : 3,50 – 3,90

結果：活性測試 : 103% ?背景 : 0,006? 顏色 : 干凈? pH : 3,75

儲存：室溫避光可短期儲存，2-8℃避光可長(cháng)期儲存

Cat. no.: 4800

Lot. no.: 170601

Date of manufacture: Jun. 2017

Exp. Date: Jun. 2021 (When stored at 2-8oC in an unopened bottle).

Analysis:The substrate is analyzed for consistent activity, background, colour and pH (only the procedure for activity and background is described in the COA). Background: 100μl substrate is pipetted into eight wells of a standard microtiter plate, followed by addition of 100μl 0.2 M sulphuric acid. The background absorbance is read at 450 nm. Activity: The activity is determined in a 2-layer ELISA. First layer is a standard human serum dilution. Second layer is rabbit anti human IgG-HRP. 100μl substrate is added to each well and incubated for 13 minutes. The reaction is stopped by addition of 100μl 0.2 M sulphuric acid. The absorbance is read at 450 nm. The activity is compared to a standard TMB solution with known activity。

Specifications:：Activity: 90 - 110% ?Background: Abs. < 0.025? Colour: Clear solution with no distinct colour? pH : 3,50 – 3,90

Results:：Activity test : 103%? ?Background : 0,006? Colour : Clear pH : 3,75

Storage:：Short term storage: Ambient; protected from light. Long term storage: 2-8℃; protected from light

Performed by: Date: 09-06-2017? Sign. : Bo Johansson

Approved by: Date: 09-06-2017? Sign. : Dorte ?rn?s

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