[KO Validated] Anti-IDH1 Rabbit Polyclonal Antibody for WB, IF/ICC, IP, ELISA - O75874

Core Product Specifications and Parameters

Biological Background: IDH1 Function and Localization

• IDH1 encodes the cytoplasmic NADP-dependent isocitrate dehydrogenase (IDH1), also known as IDPc or oxalosuccinate decarboxylase, which catalyzes the oxidative decarboxylation of isocitrate to α-ketoglutarate with concomitant production of NADPH (PubMed:10521434, PubMed:19935646).
  Related references: PMID:10521434 PMID:19935646
• The enzyme requires divalent metal ions such as Mg²⁺ or Mn²⁺ as cofactors and belongs to the isocitrate/isopropylmalate dehydrogenase family, with key roles in the tricarboxylic acid cycle and cellular redox homeostasis.
• IDH1 is predominantly localized in the cytoplasm and cytosol, and is also found in peroxisomes, where it may supply NADPH for peroxisomal reactions such as phytanoyl-CoA dioxygenase activity.
• By producing NADPH, IDH1 supports reductive biosynthesis, including lipid synthesis, and helps maintain the cellular reducing environment against oxidative stress.
• The protein is subject to post-translational modifications, including acetylation and phosphorylation, which may modulate its enzymatic activity and subcellular distribution.
• IDH1 may function as a corneal epithelial crystallin, potentially contributing to the maintenance of corneal transparency (By similarity).
• The calculated molecular weight of the mature IDH1 monomer is approximately 46.7 kDa, consistent with the observed band around 47 kDa in Western blotting.

Experimental Guidance and Technical Tips

• The immunogen encompasses the full-length human IDH1 protein (aa 1–414), suggesting the antibody can recognize both native and denatured conformations. For Western blotting, an expected band near 47 kDa should be observed; validate using appropriate positive and negative controls, as this antibody is KO-validated.
• In immunoprecipitation (IP), use a standard protocol with cell lysates from IDH1-expressing tissues or cell lines. Confirm capture by Western blotting with a different antibody or by mass spectrometry.
• For immunofluorescence/immunocytochemistry (IF/ICC), anticipate a predominantly cytoplasmic staining pattern with possible peroxisomal puncta. Include appropriate fixation and permeabilization conditions, and verify specificity using IDH1 knockout control cells if available.
• This antibody cross-reacts with mouse and rat IDH1, making it suitable for studies in rodent models. Always validate expression levels in the specific tissue or cell system under investigation.
• For ELISA, consider pairing with a matched detection antibody or using the antibody as both capture and detection in a sandwich format, ensuring minimal cross-reactivity with other isocitrate dehydrogenase isoforms (e.g., IDH2, IDH3).

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