TREM2 Antibodies

Triggering receptor expressed on myeloid cells 2 (TREM2)

Forms a receptor signaling complex with TYROBP and triggers activation of the immune responses in macrophages and dendritic cells. May have a role in chronic inflammations and may stimulate production of constitutive rather than inflammatory chemokines and cytokines.

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Gene Information

Human
Gene Name:	TREM2
Uniprot:	Q9NZC2
Entrez:	54209

Family

Belongs to:
No superfamily

Alternative Names

PLOSL2; TREM2; TREM-2; Trem2a; Trem2b; Trem2c; TREM-2triggering receptor expressed on myeloid cells 2a; Triggering receptor expressed on monocytes 2; triggering receptor expressed on myeloid cells 2

Molecular Weight

Mass (kDA):
25.447 kDA

Genomic Context

Human
Location:	6p21.1
Sequence:	6; NC_000006.12 (41158508..41163116, complement)

Tissue Specificity

Expressed on macrophages and dendritic cells but not on granulocytes or monocytes. In the CNS strongest expression seen in the basal ganglia, corpus callosum, medulla oblongata and spinal cord.

Subcellular Localizations

[Isoform 1]: Cell membrane; Single-pass type I membrane protein.; [Isoform 2]: Secreted.; [Isoform 3]: Secreted.

Diseases

• Polycystic Lipomembranous Osteodysplasia With Sclerosing Leukoencephalopathy
• Dementia
• Bone Cysts
• Inflammation
• Presenile Dementia
• Leukoencephalopathies
• Nerve Degeneration
• Alzheimer's Disease
• Nervousness
• Inflammatory Response
• Hereditary Diseases
• Autoimmune Reaction

• Subacute Sclerosing Panencephalitis
• Osteochondrodysplasias
• Multiple Sclerosis
• Lipodystrophy
• Osteopenia
• Sclerosis
• Chronic Inflammation

Interacting Proteins

• APOE
• APP
• PSEN1
• TYROBP

Pathways

• Phagocytosis
• Immune Response
• Inflammatory Response
• Pathogenesis
• Osteoclast Differentiation
• Bone Resorption
• Cell Activation
• Secretion
• Osteoclast Development
• Innate Immune Response
• Cytokine Production
• Chemotaxis
• Macrophage Activation

• Tissue Homeostasis
• Neutrophil Degranulation
• Myelination
• Cell Maturation
• System Development
• Translation
• Axon Guidance

PTMs

• Phosphorylation
• Biotinylation

• Acetylation
• Methylation

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/TREM2

Boster Bio - The Best Uses Of The TREM2 Marker

One of the most essential guides available from Boster Bio is its optimization guide. This guide will help you optimize your experiments to achieve the best results. Every researcher has run into difficulties during their research. Although there are a myriad of reasons for error, the right controls can eliminate many of them. Troubleshooting guides will help you find these problems and correct them. These guides can be helpful to identify problems and improve the overall effectiveness of your experiments.

QRT-PCR can be used to measure the levels of mRNA in tissues

In the world of biotechnology, molecular medicine, and microbiology, real-time reverse transcription polymerase chain reaction (qRT-PCR) has become the most commonly used method to measure tissue mRNA levels. But, it is not a standard assay and the issues associated with the variability of results are well-known. Fortunately the protocols for qRT-PCR outline the steps required to obtain reliable and reproducible results.

To utilize qRT-PCR to determine tissue RNA levels first, you must determine the reference gene. The reference gene must be stable in its expression and similar to the target gene. The ideal reference gene must have the same levels of expression in all experimental conditions, but increasing evidence suggests that gene expression levels can vary considerably under different experimental conditions. It is important to identify a reliable reference gene in your tissue.

It is crucial to take care to choose the reference genes. PPIA, GAPDH, and YWHAZ were the most stable genes used to reference each of the tissues. In addition, the gene expression levels of ACTB, GAPDH, and glyceraldehyde-3-phosphate dehydrogenase were evaluated and validated. The study's findings gave the basis for analyses of gene expression in future CR studies.

Quantitative real-time reverse transcriptionPCR (qRTPCR) is a different popular method of miRNA detection. It is sensitive enough to detect low levels of mature miRNA. This technique can detect up to 220 miRNAs and is extensively used for absolute quantification. However, it is not able to provide the capability of identifying novel miRNAs. However it has overcome these limitations to become the most sought-after method of miRNA expression analysis.

One of the biggest challenges in QRTPCR is selecting the right mRNA to be used in the experiment. The choice of the standard mRNA is dependent on the application. It is nonetheless essential to prove that the levels of RNA in a particular sample are invariant across samples. For instance, qRT-PCR has been utilized to detect the viral load in the lung tissue of mice following adjuvant treatment.

Using qRT-PCR for measuring tissue RNA levels has several benefits. It is a versatile method that is suitable for a variety of purposes. It is also believed as the most reliable instrument to study the expression of genes. Its high sensitivity permits it to identify and quantify target genes that have low levels of expression, and it can detect mRNA expression levels in a variety of samples. To prevent non-specific variations in cDNA samples, qRT-PCR uses reference genes that are expressed in all cells in an organism. Reference genes are also utilized to avoid contamination during sample manipulation or RNA extraction.

To perform qRTPCR, RNA samples from tissues were extracted using TRIzol(r) the reagent (Thermo Fischer Scientific). This eliminates contamination of DNA and then performs reverse transcription. Then, a different method was performed using SuperScript II to measure the target gene transcripts. The mRNA levels obtained were calculated and the qRT-PCR products were then analyzed to verify the reproducibility of the results.

A qRT-PCR assay typically comprises microtitre plates 96-well with a final volume of 25 milliliters. The qRTPCR assay mix contains five ml of CDNA and a universal mix. The mix is dispersed with a computerized eight-channel automatic pipette to reduce the variance between samples.

A high amount of TH expression in neuroblastoma indicates the presence of the disease. It is the most prevalent solid tumor that is extracranial in children. Maximizing NB minimal residual disease detection is crucial for determining the risk of returning malignant cells. Using qRT-PCR, researchers were able to detect tyrosine hydroxylase and paired-like homeobox 2b expression levels in neuroblastoma-infected testic tissue.

To diagnose NB, qRT-PCR is a highly sensitive technique to detect NB in bone blood, peripheral blood, and hematopoietic stem cells. The analysis of NB the mRNA isn't conclusive because it doesn't identify the extent of invasive potential and vitality. The results are also not sufficient to determine the success of a therapy or a cure for this disease.

Northern blot analysis (or Northern Blot Analysis) is the first method of analyzing the expression of mRNA. It allows visualization of the size of the mRNA as well as providing details about its structure. This method has a disadvantage that it doesn't allow for precise data interpretation. Because it does not have size fractionation, it is not able to distinguish the target mRNA from other sequences. It isn't precise enough to distinguish between nonspecific and target hybridization.

qRT-PCR to determine tissue protein levels

One of the most effective methods for determining the level of tissue protein levels is quantitative reverse transcription polymerase chain reaction (qRT-PCR). It is a method to assess gene expression levels in biological samples or in tissue samples. Utilizing qRTPCR as an approach to study gene expression, can be challenging due to specimen-related issues such as variability in RNA templates quality and quantity and normalization of the sample. These challenges require to change the methods used to analyze qRTPCR using small amounts of mRNA. This is especially true when working in cell groups during dissection. This article outlines the rationale for qRT-PCR analysis in tissue samples.

Because they reflect stable gene expression across tissues, it is essential to utilize housekeeping genes in qRTPCR. While ACTB shows stable gene expression in prostate cancer cells but the same expression does not match in tissue samples from ex vivo. It is possible that ACTB may not be the most stable endogenous control gene, or that its expression level will vary depending on the state of the cancer and the method of acquisition.

Single-plex reactions don't require different primer/probe sets or different dyes for the reporter, but they might require the use of a VIC labeled control gene or another endogenous control reference gene. However the majority of single-plex reactions are performed with primer-limited primer/probe sets. The paired samples should contain the same amount of cells, laser shots and total RNA. Once paired samples are analyzed using qRT-PCR, it's possible to compare the results of the experiments with other methods.

Total RNA can be extracted from tissues microdissected for qRT-PCR and stored at -80degC. The RNA is examined in two steps: first, the extraction of the total RNA, and then, the amplification of the probes and primers. In both of these steps the use of random hexamers to minimize the effects of DNA degradation. Random hexamers provide greater cDNA coverage for each individual gene.

The Cq values for the RT-qPCR experiments that use SF-OCT as well as RNL tissues are similar to those observed using PCR Endpoint. However, the Cq values of SF samples were higher than those of SF and FFPE tissues. Further studies should assess the integrity of RNA from tissues by comparing RNA samples from different kinds of samples. The qRTPCR method cannot detect RNA when tissue RNA is unstable prior to the amplification process.

When performing qRT-PCRtests, an air hood that is not leaking is necessary for the experiment. A clean environment is essential for pipettes tubes, pens, and vials for samples. In a separate hood for dead air, it is recommended to use a vortex for mixing the source vials. Before opening source vials, they should be mixed before being spun down. Before performing qRT-PCR, a positive control should be carried out for each primer.

Before performing qRT-PCR, is essential to stain samples with the specific reagent. H&E is the most popular method for visualizing. However there are methyl-green stains that will not interfere with qRTPCR. The three stains were evaluated before microdissection, and there was no statistically significant difference between the staining techniques.

KAPA PROBE FAST One-Step qRT-PCR Master Kit Universal Kit is a straightforward and reliable method for performing real-time PCR. This kit contains both ROX High Reference Dyes and ROX Low Reference Dyes. It also includes dUTP. It also includes the ABScript II One-Step RT-PCR Probe kit, as well as the KAPA PROBE FAST qRTPCR Master Mix Universal System.