Thermo Fisher Scientific DPC4 Recombinant Rabbit Monoclonal Antibody (ZR281), RAbMono

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

Ready-to-use 150-200 µL

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Product Specifications

Host/Isotype

Rabbit / IgG

Class

Recombinant Monoclonal

Type

Antibody

Clone

ZR281

Immunogen

Full-length human SMAD4 protein if (typeof window.$mangular === undefined || !window.$mangular) { window.$mangular = {}; } $mangular.antigenJson = \[{targetFamily:SMAD4,uniProtId:Q13485-1,ncbiNodeId:9606,antigenRange:1-552,antigenLength:552,antigenImageFileName:Z2596RP_SMAD4_Q13485-1_Rabbit.svg,antigenImageFileNamePDP:Z2596RP_SMAD4_Q13485-1_Rabbit_PDP.jpeg,sortOrder:1}\]; $mangular.isB2BCMGT = false; $mangular.isEpitopesModalImageMultiSizeEnabled = true;

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Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Storage conditions

2-8°C

Shipping conditions

Wet ice

Product Specific Information

This product is diluted and in a ready-to-use formulation.

A recommended positive control tissue for this product is Pancreas non-tumor cells, however positive controls are not limited to this tissue type.

The primary antibody is intended for laboratory professional use in the detection of the corresponding protein in formalin-fixed, paraffin-embedded tissue stained in manual qualitative immunohistochemistry (IHC) testing. This antibody is intended to be used after the primary diagnosis of tumor has been made by conventional histopathology using non-immunological histochemical stains.

SMAD4, also called SMAD family member 4, or DPC4 (Deleted in Pancreatic Cancer-4) is a highly conserved protein present in all human tissues. Signaling from the ligand-activated membrane receptor serine/threonine kinases to nuclear targets is mediated by DPC4. Upon ligand binding, the receptors of the TGF-β family phosphorylate SMAD proteins (SMAD1 and SMAD2). These proteins then move into the nucleus, where they activate transcription. To carry out this function, the receptor activated SMAD1 and 2 require association with DPC4, also known as SMAD4. SMAD4/DPC4 is also implicated as a tumor suppressor, since it is inactivated in more than half of pancreatic carcinomas and to a lesser extent in a variety of other cancers. The lack of SMAD4 expression is present in approximately 80% of cases of pancreatic adenocarcinoma, but rarely in endometrial (0%), colorectal (0%), ovarian (3%), lung (0%), breast (2%) adenocarcinomas, and malignant melanoma (4%). SMAD4 is an important marker for confirming a diagnosis of pancreatic adenocarcinoma.

Antibody is used with formalin-fixed and paraffin-embedded sections. Pretreatment of deparaffinized tissue with heat-induced epitope retrieval or enzymatic retrieval is recommended. In general, immunohistochemical (IHC) staining techniques allow for the visualization of antigens via the sequential application of a specific antibody to the antigen (primary antibody), a secondary antibody to the primary antibody (link antibody), an enzyme complex and a chromogenic substrate with interposed washing steps. The enzymatic activation of the chromogen results in a visible reaction product at the antigen site. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with a particular antigen.

A positive tissue control must be run with every staining procedure performed. This tissue may contain both positive and negative staining cells or tissue components and serve as both the positive and negative control tissue. External Positive control materials should be fresh autopsy/biopsy/surgical specimens fixed, processed and embedded as soon as possible in the same manner as the patient sample (s). Positive tissue controls are indicative of correctly prepared tissues and proper staining methods. The tissues used for the external positive control materials should be selected from the patient specimens with well-characterized low levels of the positive target activity that gives weak positive staining. The low level of positivity for external positive controls is designed to ensure detection of subtle changes in the primary antibody sensitivity from instability or problems with the staining methodology. A tissue with weak positive staining is more suitable for optimal quality control and for detecting minor levels of reagent degradation.

Internal or external negative control tissue may be used depending on the guidelines and policies that govern the organization to which the end user belongs to. The variety of cell types present in many tissue sections offers internal negative control sites, but this should be verified by the user. The components that do not stain should demonstrate the absence of specific staining, and provide an indication of non-specific background staining. If specific staining occurs in the negative tissue control sites, results with the patient specimens must be considered invalid.

Target Information

Smad4 is encoded by the Smad4 gene that is located on chromosome 18 in humans. Smad4 is also known as Smad family member 4, Sma- and Mad-related protein 4 or Mothers against decapentaplegic (MAD) homolog 4. Smad4 contains N-terminal MH1 (MAD homology 1) and C-terminal MH2 (MAD homology 2) globular domains that are involved in DNA binding and protein interactions respectively. Binding of the TGF-beta superfamily of ligands that includes Transforming Growth Factor -beta (TGF-beta) and bone morphogenetic protein (BMP) to its cognate receptor allows phosphorylation of Smad 1, 2, 3, 5 and 8 (R-Smad, Receptor Smad). This signals for heterotrimerization with Smad4 (co-Smad, co-mediator Smad) and translocation of the complex to the nucleus. Inhibitory or antagonistic Smad (I-Smad) that includes Smad 6 and 7, interact with activated R-Smads and attenuate the signaling pathway. Smad4 acts as a tumor suppressor protein by transcriptionally regulating its target genes such as Cyclin D1 (downregulation) and collagen (upregulation) that inhibit cell proliferation. Dephosphorylation regulates nuclear export and nucleocytoplasmic dynamics of Smads.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.