Scientific Data

Western Blot

Detection of Human, Mouse, and Rat Isocitrate Dehydrogenase 1/IDH1 antibody by Western Blot.

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Detection of Human, Mouse, and Rat Isocitrate Dehydrogenase 1/IDH1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, NIH-3T3 mouse embryonic fibroblast cell line, and Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Isocitrate Dehydrogenase 1/IDH1 at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

Isocitrate Dehydrogenase 1/IDH1 antibody in SK-BR-3 Human Cell Line by Immunocytochemistry (ICC).

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Isocitrate Dehydrogenase 1/IDH1 in SK‑BR‑3 Human Cell Line. Isocitrate Dehydrogenase 1/IDH1 was detected in immersion fixed SK-BR-3 human breast cancer cell line using Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

Isocitrate Dehydrogenase 1/IDH1 antibody in Human Brain by Immunohistochemistry (IHC-P).

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Isocitrate Dehydrogenase 1/IDH1 in Human Brain. Isocitrate Dehydrogenase 1/IDH1 was detected in immersion fixed paraffin-embedded sections of human brain (cortex) using Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to astrocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunohistochemistry

Isocitrate Dehydrogenase 1/IDH1 antibody in Rat Brain by Immunohistochemistry (IHC-Fr).

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Isocitrate Dehydrogenase 1/IDH1 in Rat Brain. Isocitrate Dehydrogenase 1/IDH1 was detected in perfusion fixed frozen sections of rat brain using Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to glial cell cytoplasm. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Knockout Validated

Western Blot Shows Human Isocitrate Dehydrogenase 1/IDH1 Antibody Specificity by Using Knockout Cell Line.

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Western Blot Shows Human Isocitrate Dehydrogenase 1/IDH1 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Isocitrate Dehydrogenase 1/IDH1 knockout HeLa cell line (KO). PVDF membrane was probed with 0.25 µg/mL of Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for Isocitrate Dehydrogenase 1/IDH1 at approximately 46 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western

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Detection of Human Isocitrate Dehydrogenase 1/IDH1 by Simple Western^TM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Isocitrate Dehydrogenase 1/IDH1 at approximately 51 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Isocitrate Dehydrogenase 1/IDH1 Monoclonal Antibody (Catalog # MAB7049). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Human Isocitrate Dehydrogenase 1/IDH1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Isocitrate Dehydrogenase 1/IDH1 in ELISAs and Western blots.

Source

Monoclonal Mouse IgG₁ Clone # 843219

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human Isocitrate Dehydrogenase 1/IDH1
Ser2-Leu414
Accession # O75874

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.25 µg/mL

See below

Simple Western

10 µg/mL

HepG2 human hepatocellular carcinoma cell line

Immunohistochemistry

8-25 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

Knockout Validated

Isocitrate Dehydrogenase 1/IDH1 is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in Isocitrate Dehydrogenase 1/IDH1 knockout HeLa cell line.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

hIDH1 MAb (Cl 843219 (100 ug)