MDH1 Antibody (JE35-03)
Novus Biologicals | Catalog # NBP3-32582
Key Product Details
Validated by
Species Reactivity
Applications
Label
Antibody Source
Product Specifications
Immunogen
Localization
Clonality
Host
Isotype
Theoretical MW
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Scientific Data Images for MDH1 Antibody (JE35-03)
Western Blot Shows MDH1 Specificity Using Knockout Cell Line.
Western blot shows lysates of HAP1 parental cell line and MDH1 knockout HAP1 cell line (KO). Nitrocellulose membrane was probed with MDH1 Antibody (JE35-03) (Catalog # NBP3-32582) followed by HRP-conjugated secondary antibody. A specific band was detected for MDH1 at approximately 36.4 kDa (as indicated) in the parental HAP1 cell line, but is not detectable in knockout HAP1 cell line. Primary antibody dilution used: 1/1000. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.Detection of MDH1 by Immunoprecipitation.
HAP1 cell line lysates were prepared and immunoprecipitation was performed using 2.0 μg of MDH1 Antibody (JE35-03) (Catalog # NBP3-32582) pre-coupled to Dynabeads Protein A. Immunoprecipitated MDH1 was detected in Western Blot with a rabbit MDH1 antibody used at 1/10000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).MDH1 Specificity is Shown by Immunocytochemistry in Knockout Cell Line.
HAP1 parental cell line WT and MDH1 HAP1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. Cells were stained with MDH1 Antibody (JE35-03) (Catalog # NBP3-32582) followed by incubation with an Alexa-fluor 555 conjugated secondary antibody (upper panel). DAPI-only counterstained cells shown on a lower panel. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. Primary antibody dilution used: 1/500. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).Western Blot: MDH1 Antibody (JE35-03) [NBP3-32582]
Western Blot: MDH1 Antibody (JE35-03) [NBP3-32582] - Western blot analysis of MDH1 on different lysates with Rabbit anti-MDH1 antibody (NBP3-32582) at 1/1,000 dilution.Lane 1: Jurkat cell lysate (20 ug/Lane)
Lane 2: HepG2 cell lysate (20 ug/Lane)
Lane 3: U-937 cell lysate (20 ug/Lane)
Lane 4: HL-60 cell lysate (20 ug/Lane)
Lane 5: NIH/3T3 cell lysate (20 ug/Lane)
Lane 6: Rat kidney tissue lysate (40 ug/Lane)
Predicted band size: 36 kDa
Observed band size: 35 kDa
Exposure time: 6 seconds; ECL;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32582) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry/ Immunofluorescence: MDH1 Antibody (JE35-03) [NBP3-32582]
Immunocytochemistry/ Immunofluorescence: MDH1 Antibody (JE35-03) [NBP3-32582] - Immunocytochemistry analysis of U-937 cells labeling MDH1 with Rabbit anti-MDH1 antibody (NBP3-32582) at 1/100 dilution.Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MDH1 antibody (NBP3-32582) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
Flow Cytometry: MDH1 Antibody (JE35-03) [NBP3-32582]
Flow Cytometry: MDH1 Antibody (JE35-03) [NBP3-32582] - Flow cytometric analysis of U-937 cells labeling MDH1.Cells were fixed and permeabilized. Then stained with the primary antibody (NBP3-32582, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Applications for MDH1 Antibody (JE35-03)
Flow Cytometry
Immunocytochemistry/ Immunofluorescence
Immunoprecipitation
Knockout Validated
Western Blot
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Formulation, Preparation, and Storage
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Background: MDH1
Long Name
Alternate Names
Gene Symbol
Additional MDH1 Products
Product Documents for MDH1 Antibody (JE35-03)
Certificate of Analysis
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Product Specific Notices for MDH1 Antibody (JE35-03)
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunoprecipitation Protocol
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Liperfluo
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars