miRCURY LNA™ microRNA ISH Optimization Kits (FFPE)에는 microRNA in situ hybridization (ISH) 실험을 setup 혹은 최적화에 필요한 모든 시약이 포함되어 있다. 본 제품에는 double DIG*-labeled miRCURY LNA™ microRNA Detection Probe와 ISH buffer와 그외 시약들을 포함하고 있으며, 각 스텝을 쉽게 진행할 수 있도록 매뉴얼도 같이 제공된다. 포함되어 있는 혹은 별도로 구매가능한 microRNA ISH Buffer Set (FFPE)는 miRCURY LNA™ microRNA Detection Probe사용에 최적화 되어 있는 고성능의 formamide-free in situ hybridization buffer이다.

The convenient way of setting up and optimizing microRNA in situ hybridization experiments in the lab
Superior sensitivity and specificity - Double DIG*-labeled miRCURY LNA™ microRNA Detection Probes and reagents specifically adapted to the kit
Very Robust - Advantageous for both high throughput and individual microRNA localization studies
Fast and easy - One-day microRNA ISH protocol
Highly flexible - No advanced instruments needed
Validated in a wide range of tissues - Ideal for use with clinical and experimental FFPE samples
ISH buffer and Proteinase K optimized for use with miRCURY LNA™ microRNA Detection Probes : Formamide-free

제품 설명

Features
A miRCURY LNA™ microRNA ISH Optimization Kit (FFPE)은 formalin-fixed paraffin embedded (FFPE) tissue 샘플의 microRNA in situ hybridizations (ISH)를 setup 혹은 최적화하기 위해 사용하는 제품이다.

(5’ and 3’) DIG*-labeled miRCURY LNA™ microRNA Detection Probe 기술을 이용하여 매우 민감하고, 특이적으로 microRNA를 검출할 수 있다. 또한 이 kit은 FFPE tissue sections에 LNA probe를 적용할 수 있는 reagent들을 제공한다.

그림1. Principle of the miRCURY LNA™ microRNA ISH Optimization Kit (FFPE). First, the tissue is “opened” using Protease K. In the hybridization step, the DIG-labeled LNA™ probe binds specifically to its target microRNA. Alkaline phosphatase (AP)-conjugated anti-DIG antibodies are then added. This step is followed by NBT-BCIP development and optional counter-staining with Nuclear Red.

함께 제공되는 instruction manual에는 ISH 각 스텝을 자세하게 설명해 두었고, 성공적인 실험을 위한 tip과 추천사항을 명시하였다. 또한 이 과정은 빠르고, 문제 없는 ISH 분석을 통해 one-day protocol이 가능하게 하였다.

Applications
본 제품은 cellular 혹은 sub-cellular microRNA의 localization 연구 및 위치에 따른 RNA 발현 결정 등을 포함한 수많은 연구에 적용할 수 있다. 또한 실험과정의 최적화는 high throughput ISH 분석과 각각의 microRNA의 localization 연구 모두에 사용이 가능하도록 하였다.

Coverage
8개의 다른 miRCURY LNA™ microRNA ISH Optimization Kit이 공급된다. 각각의 제품에는 positive, negative control probes, , hybridization buffer, Proteinase K가 포함되어 있다. 또한 각 제품에 필요한 unique tissue-specific miRCURY LNA™ microRNA Detection Probe도 같이 포함된다.

Each kit includes:

Unique microRNA LNA™ probe (double DIG*-labeled, kit specific)
Scrambled LNA™ probe (double DIG*-labeled, negative control)
U6 LNA™ probe (5’ DIG*-labeled, positive control)
Hybridization buffer (2x, formamide-free)
Proteinase K

Unique microRNA LNA™ probe는 다양한 조직에서 검증되었으므로, ISH 실험 세팅에 최적이다 (조직 종류에 따른 kit의 선택은 표1을 참조). 표 2에는 각각의 제품으로 제공되는target microRNA가 발현되는 cell을 표기하였다.

	Kit 1	Kit 2	Kit 3	Kit 4	Kit 5	Kit 7	Kit 8	Kit 9
Brain				yes
Eye				yes	yes
Muscle	yes				yes
Lung					yes	yes
Kidney					yes
Liver			yes		yes
Colon					yes	yes		yes
Cervix							yes
Heart	yes				yes	yes
Mammary Gland					yes		yes
Lung cancer		yes			yes	yes	yes
Colorectal cancer		yes			yes	yes
Breast cancer		yes			yes	yes	yes
Kidney cancer		yes			yes	yes
Cervix cancer		yes			yes	yes	yes
Testis cancer					yes	yes
Esophagus cancer								yes
Cell entity	myocyte	varies	hepatocyte	neuron	endothelial	smooth muscle	basal cells	granulocyte

표1. Choosing the appropriate miRCURY LNA™ microRNA ISH Optimization Kit. The table indicates the tissue(s) in which each of the kits has been validated.

Product	Unique probe	Examples of miR-expressing cells
Kit 1	miR-1	Skeletal muscle cells, myocytes, myocardiocytes
Kit 2	miR-21	Cancer-associated fibroblasts, some immune cells
Kit 3	miR-122	Hepatocytes
Kit 4	miR-124	Neurons
Kit 5	miR-126	Endothelial cells
Kit 7	miR-145	Smooth muscle, vascular smooth muscle (pericytes), myoepithelial cells
Kit 8	miR-205	Basal keratinocytes, myoepithelium, SCC cancer cells
Kit 9	miR-223	Myeloid, granulocytic, and monocytic compartments of the hematopoietic system

표2. Description of the kits.

Exiqon’s miRCURY LNA™ microRNA ISH Buffer Set (FFPE) includes:

ISH buffer (2x, 25 ml)
Proteinase K (12 mg, lyophilized)

Note
microRNA in situ hybridization 시작할 때에는 miRCURY LNA™ microRNA ISH Optimization Kit 사용을 추천한다. 이 제품은 ISH buffer, validated control probes, easy-to-follow manual을 포함하고 있다.

*Licensed from Roche Diagnostics GmbH

상세설명

Code	제품명	상세설명
90001	microRNA ISH Optimization Kit 1 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 1 (miR-1), controls, buffer and ProtK (400 slides)
90002	microRNA ISH Optimization Kit 2 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 2 (miR-21), controls, buffer and ProtK (400 slides)
90003	microRNA ISH Optimization Kit 3 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 3 (miR-122), controls, buffer and ProtK (400 slides)
90004	microRNA ISH Optimization Kit 4 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 4 (miR-124), controls, buffer and ProtK (400 slides)
90005	microRNA ISH Optimization Kit 5 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 5 (miR-126), controls, buffer and ProtK (400 slides)
90007	microRNA ISH Optimization Kit 7 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 7 (miR-145), controls, buffer and ProtK (400 slides)
90008	microRNA ISH Optimization Kit 8 (FFPE)	miRCURY LNA™ microRNA Detection (FFPE), Optimization Kit 8 (miR-205), controls, buffer and ProtK (400 slides)
90009	microRNA ISH Optimization Kit 9 (FFPE)	miRCURY LNA; microRNA Detection (FFPE), Optimization Kit 9 (miR-223), controls, buffer and ProtK (400 slides)
90010	microRNA ISH Buffer and Controls Kit	miRCURY LNA; microRNA Detection, control probes, buffer and ProtK (400 slides)
90000	microRNA ISH Buffer Set (FFPE)	miRCURY LNA™ microRNA Detection, microRNA ISH Buffer (FFPE) and ProtK, (1000 slides)