Protocol
Mouse anti-Human IDH1 R132H Monoclonal Antibody | anti-IDH1 R132H antibody
IDH1 R132H
Reactivity
Human
Applications
Immunohistochemistry
Synonyms
IDH1 R132H, Antibody; IDH1 R132H; anti-IDH1 R132H antibody
Host
Mouse
Reactivity
Human
Clonality
Monoclonal
Isotype
IgG1
Clone Number
IHC132
Form/Format
Prediluted
Applicable Applications for anti-IDH1 R132H antibody
IHC (Immunohistochemistry)
Immunogen
Synthetic peptide corresponding to the human IDH1 surrounding residual 132.
Localization
Cytoplasmic
Control
Glioma, Glioblastoma, Astrocytoma with IDH1 R132H Mutation
Presentation
ADHL 1 R132H is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.
Specimen Preparation
Paraffin sections:
The antibody can be used on formalin-fixed paraffin-embedded (FFPE) tissue sections. Ensure tissue undergoes appropriate fixation for best results. Pre-treatment of tissues with heat-induced epitope retrieval (HIER) is recommended using ImmunoDNA Retriever with Citrate, ImmunoDNA Retriever with EDTA or ImmunoDNA Digestor. Tissue should remain hydrated via use of Immuno/DNA Washer solutions.
Frozen sections and cell preparations:
The antibody can be used for labeling acetone-fixed frozen sections and acetone-fixed cell preparations.
The antibody can be used on formalin-fixed paraffin-embedded (FFPE) tissue sections. Ensure tissue undergoes appropriate fixation for best results. Pre-treatment of tissues with heat-induced epitope retrieval (HIER) is recommended using ImmunoDNA Retriever with Citrate, ImmunoDNA Retriever with EDTA or ImmunoDNA Digestor. Tissue should remain hydrated via use of Immuno/DNA Washer solutions.
Frozen sections and cell preparations:
The antibody can be used for labeling acetone-fixed frozen sections and acetone-fixed cell preparations.
IHC Protocol
1. Cut and mount 3-5 micron formalin-fixed paraffin-embedded tissues on positively charged slides such as Hydrophilic Plus Slides.
2. Air dry for 2 hours at 58°C.
3. Deparaffinize and rehydrate tissues.
4. Subject tissues to heat induced epitope retrieval (HIER) using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate or EDTA.
5. Any of three heating methods may be used:
a. TintoRetriever Pressure Cooker or Equivalent
Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, and place on trivet in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high. Incubate for 15 minutes. Open and immediately transfer slides to room temperature.
b. TintoRetriever PT Module or Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA at 95°-99°C. Incubate for 30-60 minutes.
c. Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
8. Wash slides with ImmunoDNA washer or DI water.
9. Continue IHC staining protocol. Wash slides between each step with ImmunoDNA washer solution.
2. Air dry for 2 hours at 58°C.
3. Deparaffinize and rehydrate tissues.
4. Subject tissues to heat induced epitope retrieval (HIER) using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate or EDTA.
5. Any of three heating methods may be used:
a. TintoRetriever Pressure Cooker or Equivalent
Place tissues/slides in a staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA, and place on trivet in the pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high. Incubate for 15 minutes. Open and immediately transfer slides to room temperature.
b. TintoRetriever PT Module or Water Bath Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA at 95°-99°C. Incubate for 30-60 minutes.
c. Conventional Steamer Method
Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
8. Wash slides with ImmunoDNA washer or DI water.
9. Continue IHC staining protocol. Wash slides between each step with ImmunoDNA washer solution.
Mounting Protocols
For detailed instructions using biodegradable permanent mounting media such as XyGreen PermaMounter or organic solvent based resin such as PermaMounter refer to P10174 or P10097.
Product Limitations
Due to inherent variability present in immunohistochemical procedures (including fixation time of tissues, dilution factor of antibody, retrieval method utilized, and incubation time), optimal performance should be established through the use of positive and negative controls.
Preparation and Storage
Store at 2-8°C (Control Slides: Store at 20-25°C)
Protocol
Protocol
IHC (Immunohistochemistry)
(IHC of IDHL 1 R132H on a FFPE Glioma Tissue)
IHC (Immunohistochemistry)
(IHC of IDHL 1 R132H on a FFPE Glioma Tissue)
Related Product Information for anti-IDH1 R132H antibody
The IDH7 gene on chromosome band 2q33.3 encodes for isocitrate dehydrogenase 1 (IDH1), located in the cytoplasm and the peroxisomes. This enzyme catalyzes NADPH production via oxidative decarboxylation of isocitrate to alpha-ketoglutarate in the Krebs citric acid cycle. Studies have shown that IDH1 mutation is an early step in gliomagenesis and has been reported to occur in grades II and III astrocytomas, oligodendrogliomas (OG), oligoastrocytomas (OA), and secondary GBM.
Mutations involving IDH1 occur in a high proportion of diffuse gliomas, with implications on identification. About 90% involve exon 4 at codon 132, replacing amino acid arginine with histidine (R132H). Preliminary studies comparing Immunohistochemistry (IHC) with IDH1-R132H mutation-specific antibodies have shown concordance with DNA sequencing and no cross-reactivity with wild-type IDH1 or other mutant proteins.
Mutations involving IDH1 occur in a high proportion of diffuse gliomas, with implications on identification. About 90% involve exon 4 at codon 132, replacing amino acid arginine with histidine (R132H). Preliminary studies comparing Immunohistochemistry (IHC) with IDH1-R132H mutation-specific antibodies have shown concordance with DNA sequencing and no cross-reactivity with wild-type IDH1 or other mutant proteins.
Similar Products
Product Notes
The IDH1 R132H (Catalog #AAA59310) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The IDH1 R132H reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's IDH1 R132H can be used in a range of immunoassay formats including, but not limited to, IHC (Immunohistochemistry). Researchers should empirically determine the suitability of the IDH1 R132H for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "IDH1 R132H, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
If you are ready to order, navigate to Shopping Cart and get ready to checkout.