Thermo Fisher Scientific GATA-3 Recombinant Rabbit Monoclonal Antibody (ZR65), RAbMono

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:200

-

Product Specifications

Host/Isotype

Rabbit / IgG

Expression System

CHO cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

ZR65

Immunogen

Full-length human GATA-3 protein

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

100 µg/mL

Storage conditions

2-8°C

Shipping conditions

Wet ice

Product Specific Information

A recommended positive control tissue for this product is Breast cancer with ER+, Urothelial carcinoma, 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.

GATA-3 is a transcription factor that in humans is encoded by the GATA3 gene. The GATA-3 transcription factor is critical for the embryonic development of various tissues as well as for inflammatory and hormonal immune responses. This 50 kDa nuclear protein regulates the development and subsequent maintenance of a variety of human tissues, including hematopoietic cells, skin, kidney, mammary gland, and the central nervous system. Among several other roles, GATA-3 involved in luminal cell differentiation in the mammary gland and appears to control a set of genes involved in the differentiation and proliferation of breast cancer. The expression of GATA-3 is associated with the expression of estrogen receptor-alpha (ER) in breast cancer. GATA-3 has been shown to be a novel marker for bladder cancer.

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

The genes for all 4 subunits of the T-cell antigen receptor (alpha, beta, gamma and delta) are controlled by distinct enhancers and their enhancer-binding proteins. Marine and Winoto (1991) identified a common TCR regulatory element by demonstrating binding of the enhancer-binding protein GATA3 to the enhancer elements of all 4 TCR genes. GATA3 had been shown in the chicken to be an enhancer-binding protein containing a zinc finger domain. GATA3 mRNA was demonstrated by Northern blot analysis in T cells but not in B cells or macrophages. GATA3 is abundantly expressed in the T-lymphocyte lineage and is thought to participate in T-cell receptor gene activation through binding to enhancers. Labastie et al. (1994) cloned the human gene and the 5-prime end of the mouse gene. The human gene comprises 6 exons distributed over 17 kb of DNA. Its 2 zinc fingers are encoded by 2 separate exons highly conserved with those of GATA1, but no other structural homologies between the 2 genes could be found.

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