MASTER POLYMER PLUS ALKALINE PHOSPHATASE DETECTION SYSTEM

MASTER POLYMER PLUS ALKALINE PHOSPHATASE DETECTION SYSTEM

 Available Size : 100 test / 500 test

Kit components:
DAB Substrate Buffer (READY-TO-USE)
DAB Chromogen Concentrate (READY-TO-USE)
DAB enhancer (READY-TO-USE)

Master Polymer Plus AP is the result of the experience of a decade in systems of polymers for
immunohistochemistry. The detection system has proven greater sensitivity with an increase in the antigenantibody
binding signal. The new technology of micropolymers provides greater advantages as compared to
the conventional immunohistochemistry techniques and facilitates the detection of antigens in the different
cellular compartments (nucleus, cytoplasm, cytoplasmic membrane) due to the smaller size of the molecule.
The technical procedure used eliminates the background staining originated by the non-specific binding to
endogenous biotin molecules, as Master Polymer Plus AP is a procedure not based in the reaction
avidin/biotin.
The detection system Master Polymer Plus AP is highly sensitive, provides a low background staining and
the results obtained are higher than the ones obtained with the conventional procedures of
Streptavidin/Biotin or long polymers.
The system Master Polymer Plus AP is elaborated with the technology of micropolymers.
This system can be used with:
– Monoclonal primary antibodies obtained in mouse.
– Monoclonal and polyclonal primary antibodies obtained in rabbit.

دسته: ,

توضیحات و استفاده

Applicatin Of The Product: This Kit is developed for immunohistochemical study using monoclonal or polyclonal primary antibodies.
After incubating the selections with the unmarked primary antibody, the universal amplifier of primary antibodies is applied, followed by a complex constituted by micropolymer with universal secondary antibody and molecules of alkaline phosphatase. After the predetermined incubation times and after washing, it is developed using a specific chromogen of Alkaline Phosphatase (AP) type or the reagents of the developed kit.

Kit Components:
MAD-000230QK (500 test)                    MAD-000230QK-10 (100 test)
MAD-000230QK-B                                MAD-000230QK-B10                              Primary Antibodies / Amplifier Master / AP
                                                                                                                    Primary antibodies / amplifier Master AP

MAD-000230QK-C                                MAD-000230QK-C10                              Master Polymer / Plus AP
                                                                                                                    Polymer Master Plus / AP

MAD-001815QK-A                                MAD-001818QK-A                                 AP Substrate Buffer / Substrate AP

MAD-001815QK-B                                MAD-001818QK-B                                AP Chromogen / Concentrate
                                                                                                                   Chromogen AP / concentrated / solution
Reagents and Accessories Needed Not Provided With The Kit:
1) Reagents: Reagents for the deparaffining and antigenic recovery for each antibody, primary antibodies,
washing buffers and bi-distilled water (distilled deionised water or equivalent).
2) Accessories: Microscope, slide and coverslip for microscopes, pipettes, sample tubes, humid hatchery
chamber and media mounting.

Recommendations For Use:

Sample Preparation (for paraffin-embedded tissues)
The sample can experiment an antigenic denaturalization if it is subjected to a prolonged fixation. Therefore,
and in order to obtain an optimal fixation with the tissue maintaining its antigenic activity, it is
recommended the fixation with 10% buffered formalin for 24-48 hours.

Sections Preparation (for paraffin-embedded tissues)
The sections are cut at 3μm and placed on the slides. If there is a need to do more treatments as antigenic
recovery, through heat or enzymatic treatment, the crystal slides must be covered with a sticker for tissue
sections as 0.02% poly-L-lysina or silane.
It is recommended to use a tissue sample with positive immunoreactivity and another one negative, or
substitute the primary antibody with washing buffer or normal serum and process them the same way as
the template sample for a correct interpretation of the staining results.

B. Staining Procedure:

1. Incubation with the primary antibody
– Covering the template tissue section following the recommendations provided by the manufacturer for the
use of this antibody.
– Clearing in TBS 3 times for 5 minutes.

2. Incubation with the amplifier of primary antibodies Master AP:
– Applying two drops (100 μL) of the Amplifier (Master Polymer Plus AP) to each one of the samples to
completely cover the sections. Incubating at room temperature for 10 minutes.
– Clearing in TBS 3 times for 5 minutes

3. Incubating with the Polymer Master Plus AP:
– Applying two drops (100 μL) of the micropolymer (Master Polymer Plus AP) to each one of the samples to
completely cover the sections. Incubating at room temperature for 15 minutes.
Note: The micropolymer is sensitive to the light. Avoid the unnecessary exposure to light and store in an
opaque vial or container.
– Clearing in TBS 3 times for 5 minutes.

4. Developing of the immunostaining (preparation and incubation with substrate/chromogen)
The mix substrate/chromogen must be preparedin the moment in which it is needed or, maximum, 30 minutes earlier.
Adding 1 drop of red Chromogen AP concentrated at 2.5 ml of substrate buffer (or 2 drops at 5ml). Mix well. This solution must be safeguarded from light.
– Applying the mix substrate/chromogen to each one of the samples until completely covering the sections.
– Incubating at room temperature for 15 minutes.
– Washing with distilled water 3 times for minutes.

5. Sample contrast
– Covering the sample with haematoxylin for contract staining, for 1-2 min according to the intensity of the desired contrast.
– It is recommended not to use alcoholic solutions of contrast haematoxylin
– Washing properly with bi-distilled or deionized water. 6. Clearing and mounting
– After washing with water, drawing the slides at 50-60ºC for at least 30 minutes or 1 hour at room temperature, clearing in xylene and install with permanent mounting medium.

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