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Merck

DUO92005

Duolink® In Situ PLA® Probe Anti-Rabbit MINUS

Duolink® In Situ PLA® Probe Anti-Rabbit MINUS
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30 REACTIONS

$442.00

100 REACTIONS

$762.00

$442.00


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About This Item

UNSPSC Code:
12352203
NACRES:
NA.32

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biological source

donkey (polyclonal)

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody product type

primary antibodies

product line

Duolink®

species reactivity

rabbit

technique(s)

immunofluorescence: suitable
proximity ligation assay: suitable

suitability

suitable for brightfield
suitable for fluorescence

shipped in

wet ice

storage temp.

2-8°C

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1 of 4

This Item
DUO92002DUO92001DUO92006
species reactivity

rabbit

species reactivity

rabbit

species reactivity

mouse

species reactivity

goat

biological source

donkey (polyclonal)

biological source

donkey (polyclonal)

biological source

donkey (polyclonal)

biological source

donkey (polyclonal)

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody form

affinity purified immunoglobulin (secondary antibody)

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

storage temp.

2-8°C

shipped in

wet ice

shipped in

wet ice

shipped in

wet ice

shipped in

wet ice

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

technique(s)

immunofluorescence: suitable, proximity ligation assay: suitable

Application

Duolink®proximity ligation assay(PLA®) allows for endogenous detection of protein interactions, post translational modifications, and protein expression levels at the single molecule level in fixed cells and tissue samples.

This product can be applied to both the Duolink® In Situ Fluorescence Protocol and the Duolink® In Situ Brightfield Protocol depending on the detection reagents used.

Visit our Duolink® PLA Resource Center for information on how to run a Duolink® experiment, applications, troubleshooting, and more.

To perform a complete Duolink® PLA in situ experiment you will need two primary antibodies (PLA, IHC, ICC or IF validated) that recognize two target epitopes. Other necessary reagents include a pair of PLA probes from different species (one PLUS and one MINUS), detection reagents, wash buffers, and mounting medium. Note that the primary antibodies must come from the same species as the Duolink® PLA probes. Analysis is carried out using standard immunofluorescence assay equipment.HRP is also available for brightfield detection.
Specificity
PLA probe anti-Rabbit reacts with whole molecule rabbit IgG and the light chains of other rabbit immunoglobulin?s. The PLA probe anti-Rabbit has minimal cross reactivity with bovine, chicken, goat, guinea pig, Syrian hamster, horse, human, mouse, rat, and sheep serum proteins. A PLUS probe of a different species must be used simultaneously with this product. See our Product Selection Guide for more information.

Application Note
Two primary antibodies raised in different species 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. Duolink® PLA in situ reagents are suitable for use on fixed cells, cytospin cells, cells grown on slide, formalin-fixed, paraffin embedded (FFPE), or tissue (fresh or frozen). No minimum number of cells is required.

Let us do the work for you, learn more about our Custom Service Program to accelerate your Duolink® projects

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Features and Benefits

  • No overexpression or genetic manipulation required
  • High specificity (fewer false positives)
  • Single molecule sensitivity due to rolling circle amplification
  • Relative quantification possible
  • No special equipment needed
  • Quicker and simpler than FRET
  • Increased accuracy compared to co-IP
  • Publication-ready results

Other Notes

This product is comprised of the following:
  • 5x PLA Probe Anti-Rabbit MINUS - Donkey anti-rabbit secondary antibody conjugated to oligonucleotide MINUS
  • 1x Blocking Solution - Reagent for blocking of the sample
  • 1x Antibody Diluent - For dilution of PLA probes and primary antibodies
See datasheet for more information.

Legal Information

Duolink is a registered trademark of Merck KGaA, Darmstadt, Germany
PLA is a registered trademark of Merck KGaA, Darmstadt, Germany

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Storage Class Code

10 - Combustible liquids


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Roberto Di Maio et al.
Science translational medicine, 10(451) (2018-07-27)
Missense mutations in leucine-rich repeat kinase 2 (LRRK2) cause familial Parkinson's disease (PD). However, a potential role of wild-type LRRK2 in idiopathic PD (iPD) remains unclear. Here, we developed proximity ligation assays to assess Ser1292 phosphorylation of LRRK2 and, separately
Hyun Jik Lee et al.
Cell death and differentiation, 26(9), 1716-1734 (2018-11-23)
Hypoxia inducible factor 1α (HIF1α) is a master regulator leading to metabolic adaptation, an essential physiological process to maintain the survival of stem cells under hypoxia. However, it is poorly understood how HIF1α translocates into the nucleus in stem cells
Pengcheng Zhu et al.
Cancer cell, 19(3), 401-415 (2011-03-15)
Cancer is a leading cause of death worldwide. Tumor cells exploit various signaling pathways to promote their growth and metastasis. To our knowledge, the role of angiopoietin-like 4 protein (ANGPTL4) in cancer remains undefined. Here, we found that elevated ANGPTL4
M Aubele et al.
British journal of cancer, 103(5), 663-667 (2010-08-12)
Protein tyrosine kinase 6 (PTK6; breast tumour kinase) is overexpressed in up to 86% of the invasive breast cancers, and its association with the oncoprotein human epidermal growth factor receptor 2 (HER2) was shown in vitro by co-precipitation. Furthermore, expression
Kan V Lu et al.
Cancer cell, 22(1), 21-35 (2012-07-14)
Inhibition of VEGF signaling leads to a proinvasive phenotype in mouse models of glioblastoma multiforme (GBM) and in a subset of GBM patients treated with bevacizumab. Here, we demonstrate that vascular endothelial growth factor (VEGF) directly and negatively regulates tumor

Articles

Support information including tips and tricks, frequently asked questions, and basic troubleshooting.

Things to consider for preparation, setup and execution of the Duolink® assay protocol

包括建议和技巧、常见问题解答和基本故障排除的支持信息。

正确地准备、设置和执行 Duolink® PLA 实验方案需要考虑的因素。

Protocols

使用明场显微镜和Duolink® PLA试剂用于检测和定量组织和细胞样品中的个体蛋白质、蛋白质修饰和蛋白质相互作用。

本实验方案描述如何使用Duolink® PLA邻位连接试剂,通过流式细胞术检测细胞群内的单个蛋白质、蛋白修饰和蛋白相互。

Duolink® PLA reagents enable brightfield detection and quantification of proteins and interactions in tissue samples.

Related Content

Applications to detect, quantify and visualize protein-protein interactions, post-translational modifications and low expression protein detection using proximity ligation assay

利用邻位连接技术对蛋白质互作、翻译后修饰和低表达蛋白进行检测、定量和成像应用。

Questions

1–5 of 5 Questions  
  1. hello, I want to know the volume of "PLA probe anti-mouse plus (duo92001)" 30rxn.

    1 answer
    1. The 30 reaction Duolink PLUS and MINUS probe components are supplied at 240 μL.

      Helpful?

  2. What epitope on the antibody is recognized by the Duolink PLA Probes?

    1 answer
    1. The Duolink PLA Probes offered are donkey polyclonal antibodies directed to the whole molecule (heavy and light chains).  They are affinity purified.  The same polyclonal antibody is conjugated to the Plus and Minus Oligos. The Plus and Minus probes can be used together (to detect a single antibody) since they can recognize multiple sites.  The anti-mouse IgG PLA labeled antibodies may cross-react with rat but this is not consistent. Do not use this product with rat antibodies.  It is best to label anti-rat secondary antibodies with the Duolink Probemaker kit.The anti-goat IgG PLA labeled antibodies have cross-reactivity with sheep IgG and can be recommended for this application.

      Helpful?

  3. How many reactions can be performed with the Duolink® In Situ Starter Kit?

    1 answer
    1. All of the starter kits for the Duolink product line will perform 30 reactions using 40 ul per reaction.    DUO92101 - Duolink® In Situ Red Starter Kit Mouse/Rabbit    DUO92102 - Duolink® In Situ Orange Starter Kit Mouse/Rabbit    DUO92103 - Duolink® In Situ Red Starter Kit Mouse/Goat    DUO92104 - Duolink® In Situ Orange Starter Kit Mouse/Goat    DUO92105 - Duolink® In Situ Red Starter Kit Goat/Rabbit    DUO92106 - Duolink® In Situ Orange Starter Kit Goat/Rabbit

      Helpful?

  4. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  5. Has Duolink been used for detection of changes in protein expression?

    1 answer
    1. The following reference cites the use of Duolink to detect changes in protein expression in Human. Raizer et al., Cancer 2010;116:5297-305

      Helpful?

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