Thermo Fisher Scientific IDH1 Recombinant Rabbit Monoclonal Antibody (ZR250)

Applications

Tested Dilution

Publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:200

-

Product Specifications

Host/Isotype

Rabbit / IgG

Class

Recombinant Monoclonal

Type

Antibody

Clone

ZR250

Immunogen

Synthetic peptide, amino acid sequence proprietary

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 Glioma, 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.

Isocitrate dehydrogenase 1 is an enzyme that in humans is encoded by the IDH1 gene on chromosome 2. Isocitrate dehydrogenase catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate.
IDH1 mutations are heterozygous, typically involving an amino acid substitution in the active site of the enzyme in codon 132. Mutations in IDH1 (R132H) are implicated in cancer. IDH1 and its homologue IDH2 are among the most frequent mutations in diffuse gliomas including diffuse astrocytomas, anaplastic oligodendrogliomas, oligoastrocytomas, and secondary glioblastoma. Mutations in IDH1 are often the first hit in the development of diffuse gliomas, suggesting IDH1 mutations as key events in the formation of these brain tumors. Glioblastomas with a wild-type IDH1 gene have a median overall survival of only 1 year, whereas IDH1-mutated glioblastoma patients have a median overall survival of over 2 years. Tumors of various tissue types with IDH1/2 mutations show improved responses to radiation and chemotherapy.
Antibody reacts specifically with IDH1 (R132H) point mutation.

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

Isocitrate dehydrogenases such as IDH1 catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which utilizes NAD(+) as the electron acceptor and the other NADP(+). Five isocitrate dehydrogenases have been reported: three NAD(+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP(+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP(+)-dependent isozyme is a homodimer. The protein encoded by this gene is the NADP(+)-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. It contains the PTS-1 peroxisomal targeting signal sequence. The presence of this enzyme in peroxisomes suggests roles in the regeneration of NADPH for intraperoxisomal reductions, such as the conversion of 2, 4-dienoyl-CoAs to 3-enoyl-CoAs, as well as in peroxisomal reactions that consume 2-oxoglutarate, namely the alpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production. This product is specific for IDH1 (mutant R132H).

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