DUO92004

Sigma-Aldrich

Duolink® In Situ PLA® Probe Anti-Mouse MINUS

Affinity purified Donkey anti-Mouse IgG (H+L)

NACRES:
NA.32

biological source

donkey (polyclonal)

Quality Level

antibody form

affinity purified immunoglobulin (secondary antibody)

antibody product type

primary antibodies

product line

Duolink®

species reactivity

mouse

application(s)

immunofluorescence: suitable
proximity ligation assay: suitable

suitability

suitable for brightfield
suitable for fluorescence

shipped in

wet ice

storage temp.

2-8°C

General description

Duolink® In Situ PLA® Probe Anti-Mouse MINUS contains affinity purified donkey anti-mouse IgG (H+L), which reacts with whole molecule mouse IgG and also reacts with the light chains of other mouse immunoglobulins.

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-Mouse reacts with whole molecule Mouse IgG and the light chains of other Mouse immunoglobulin′s. The PLA probe anti-Mouse may cross-react with Rat antibodies, but has minimal cross reactivity with bovine, chicken, goat, guinea pig, Syrian hamster, horse, human, rabbit, 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

View full Duolink® product list

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

Components

This product is comprised of the following:
  • 5x PLA Probe Anti-Mouse MINUS - Donkey anti-mouse 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.

Preparation Note

Storage and Stability: Store the PLA Probe Anti-Mouse MINUS at 2–8°C. Do not store the diluted PLA Probe Anti-Mouse MINUS solution.

Legal Information

Duolink is a registered trademark of Sigma-Aldrich Co. LLC
PLA is a registered trademark of Sigma-Aldrich Co. LLC

RIDADR

NONH for all modes of transport

Proximal Ligation Assay (PLA) on Lung Tissue and Cultured Macrophages to Demonstrate Protein-protein Interaction.
Mendez R and Santanu B
Bio-protocol, 7(21) (2017)
Novel function of ceramide for regulation of mitochondrial ATP release in astrocytes.
Kong J N, et al.
Journal of Lipid Research, M081877-M081877 (2018)
Interaction of the Mouse Polyomavirus Capsid Proteins with Importins Is Required for Efficient Import of Viral DNA into the Cell Nucleus.
Soldatova I, et al.
Viruses, 10(4), 165-165 (2018)
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...
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