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Duolink®
기술
flow cytometry: suitable
immunofluorescence: suitable
proximity ligation assay: suitable
형광
λex 644 nm; λem 669 nm
적합성
suitable for fluorescence
배송 상태
dry ice
저장 온도
−20°C
일반 설명
Duolink® flowPLA Detection Kit contains detection oligonucleotides with a fluorophore (lex = 644 nm/lem = 669 nm).
특이성
Far Red Fluorescence Detection Reagents
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission
Use appropriate laser for λex 644 nm excitation
Use appropriate filter for λem 669 nm emission
애플리케이션
Based on proximity ligation assay (PLA), the Duolink® PLA Technology allows for endogenous detection of protein interactions, post-translational modifications, and protein expression levels at the single molecule level in fixed cells.
Duolink® flowPLA Detection Kits will enable sensitive detection of proteins, protein-protein interactions, and protein modifications within cell populations by flow cytometry. To perform a Duolink® flowPLA experiment, you will need fixed, suspended cells, two primary antibodies that specifically recognize your proteins of interest, a pair of PLA probes (one 100RXN PLUS and one 100RXN MINUS), wash buffer, and a Duolink® flowPLA Detection Kit. The flowPLA Kits are available with 5 different fluorophores: Violet, Red, Green, Orange, or FarRed. The flowPLA Kits contain all the necessary reagents to perform the amplification and detection of bound PLA probes by flow cytometry. Analysis is carried out using standard flow cytometry assay equipment. User must provide a fixed cell suspension, primary antibodies, and corresponding PLA Probes.
Follow the Duolink® PLA Flow Cytometry Protocol to use this product.
Visit our Duolink® PLA Flow Cytometry page on how to run a Duolink® flow experiment, applications, troubleshooting, and more.
Application Note
Primary antibodies are needed. Test your primary antibodies (IgG-class, mono- or polyclonal) in a standard immunofluorescence (IF), immunohistochemistry (IHC), or immunocytochemistry (ICC) assay to determine the optimal fixation, blocking, and titer conditions. Flow validated antibodies are recommended.
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects
View full Duolink® product list
Duolink® flowPLA Detection Kits will enable sensitive detection of proteins, protein-protein interactions, and protein modifications within cell populations by flow cytometry. To perform a Duolink® flowPLA experiment, you will need fixed, suspended cells, two primary antibodies that specifically recognize your proteins of interest, a pair of PLA probes (one 100RXN PLUS and one 100RXN MINUS), wash buffer, and a Duolink® flowPLA Detection Kit. The flowPLA Kits are available with 5 different fluorophores: Violet, Red, Green, Orange, or FarRed. The flowPLA Kits contain all the necessary reagents to perform the amplification and detection of bound PLA probes by flow cytometry. Analysis is carried out using standard flow cytometry assay equipment. User must provide a fixed cell suspension, primary antibodies, and corresponding PLA Probes.
Follow the Duolink® PLA Flow Cytometry Protocol to use this product.
Visit our Duolink® PLA Flow Cytometry page on how to run a Duolink® flow experiment, applications, troubleshooting, and more.
Application Note
Primary antibodies are needed. Test your primary antibodies (IgG-class, mono- or polyclonal) in a standard immunofluorescence (IF), immunohistochemistry (IHC), or immunocytochemistry (ICC) assay to determine the optimal fixation, blocking, and titer conditions. Flow validated antibodies are recommended.
Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects
View full Duolink® product list
Duolink® flowPLA Detection Kit–FarRed has been used in in situ proximity ligation assay:
- to detect interaction and complex formation between cellular retinoic acid binding protein 1 (Crabp1) and the components of rapidly accelerated fibrosarcoma (Raf) kinase- MAPK-Erk kinase (Mek) signaling pathway
- to study protein interaction in human cultured MOLT-4 cells and HeLa cells
- to visualize Beclin-1 protein interaction with 14-3-3t in neurons
- to study protein interactions in graft endothelial cells
특징 및 장점
- Analyze protein protein interactions with flow cytometry readout
- Analyze cell populations with Proximity Ligation Assay
- Increased sensitivity due to rolling circle amplification for low abundant targets
- No overexpression or genetic manipulation required
- Relative quantification possible
- Works with any flow cytometer instrumentation
- Easy to follow flexible protocol
- Publication-ready results
성분
This product is comprised of the following:
See datasheet for more information.
- 5x Detection Solution - FarRed (DUO84004)
- 5x Ligation Buffer (DUO82009)
- 5x Amplification Buffer (DUO82050)
- Ligase (1U/μL)
- Polymerase (10U/μL)
See datasheet for more information.
법적 정보
Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany
신호어
Danger
유해 및 위험 성명서
예방조치 성명서
Hazard Classifications
Resp. Sens. 1
Storage Class Code
10 - Combustible liquids
이미 열람한 고객
Tyler J Burns et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 91(2), 180-189 (2017-01-18)
To quantify visual and spatial information in single cells with a throughput of thousands of cells per second, we developed Subcellular Localization Assay (SLA). This adaptation of Proximity Ligation Assay expands the capabilities of flow cytometry to include data relating
Sofie Selmer Andersen et al.
Cytokine, 64(1), 54-57 (2013-06-04)
Many cytokine receptors are cell surface proteins that promiscuously combine to form active signalling homo- or heterodimers. Thus, receptor chain dimerization can be viewed as a direct measure of a high probability of intracellular signalling by specific cytokines. Proximity ligation
Karl-Johan Leuchowius et al.
Cytometry. Part A : the journal of the International Society for Analytical Cytology, 75(10), 833-839 (2009-08-04)
Interactions between members of the epidermal growth factor receptor (EGFR) family mediates cellular responses to ligand stimulation. Measurement of these interactions could provide important information and may prove useful as prognostic markers in malignancy. Therefore, to develop methods to study
Teng-Jian Zhou et al.
Theranostics, 7(5), 1389-1406 (2017-04-25)
Cancer stem cells (CSCs) are a small subset of malignant cells, possessing stemness, with strong tumorigenic capability, conferring resistance to therapy and leading to the relapse of nasopharyngeal carcinoma (NPC). Our previous study suggested that cyclooxygenase-2 (COX-2) would be a
Valerie Le Sage et al.
Virology, 502, 73-83 (2016-12-26)
Stress granules (SGs) are dynamic cytoplasmic aggregates of translationally silenced mRNAs that assemble in response to environmental stress. SGs appear to play an important role in antiviral innate immunity and many viruses have evolved to block or subvert SGs components
문서
Considerations for proper experimental design, preparation and execution of the Duolink® PLA for flow cytometry protocol.
General tips and tricks for proper experiment execution, aid in identifying potential problems, and provide solutions to ensure a successful Duolink® PLA experiment for flow cytometry.
Traditional flow cytometry has been limited in the ability to detect protein-protein interactions and low abundant proteins events — until now. We have combined Duolink® Proximity Ligation Assay (PLA) with flow cytometry in a convenient kit, making the analysis of protein-protein interactions with flow cytometry readouts a reality.
자사의 과학자팀은 생명 과학, 재료 과학, 화학 합성, 크로마토그래피, 분석 및 기타 많은 영역을 포함한 모든 과학 분야에 경험이 있습니다..
고객지원팀으로 연락바랍니다.