4-Hydroxy-2-hexenal Antibody: APC

• Catalog Number: orb396578
• Category: Antibodies
• Description: Mouse monoclonal antibody against 4-Hydroxy-2-hexenal conjugated to APC .
• Target: 4-Hydroxy-2-hexenal
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: IgG2b
• Conjugation: APC
• Reactivity: All
• Concentration: 1 mg/ml
• Buffer/Preservatives: 95.64mM Phosphate, 2.48mM MES and 2mM EDTA
• Purification: Protein G Purified
• Immunogen: Synthetic 4-Hydroxy-2-hexenal modified Keyhole Limpet Hemocyanin (KLH).
• Tested applications: ICC, IF, IHC
• Dilution range: WB (1:1000); ICC/IF (1:50); ELISA (1:1000); FACS (1:50); FCM (1:50)
• Application notes: A 1:1000 dilution of SMC-510 was sufficient for detection of 4-Hydroxy-2-hexenal in 0.5 µg of 4-Hydroxy-2-hexenal conjugated to BSA and 4-Hydroxy nonenal conjugated to BSA by ECL immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary Antibody.
• Clone Number: 6F10
• Storage: Conjugated antibodies should be stored according to the product label
• Alternative names: 4-Hydroxy-2-hexenal (4-HHE) antibody, 4-Hydroxy-2-
• Research Area: Cancer Biology, Cell Biology, Metabolism Research
• Note: For research use only

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-4-Hydroxy-2-hexenal Monoclonal Antibody, Clone 6F10. Tissue: Embryonic kidney epithelial cell line (HEK293). Species: Human. Fixation: 5% Formaldehyde for 5 min. Primary Antibody: Mouse Anti-4-Hydroxy-2-hexenal Monoclonal Antibody at 1:400 for 30-60 min at RT. Secondary Antibody: Goat Anti-Mouse Alexa Fluor 488 at 1:1500 for 30-60 min at RT. Counterstain: Phalloidin Alexa Fluor 633 F-Actin stain; DAPI (blue) nuclear stain at 1:250, 1:50000 for 30-60 min at RT. Magnification: 20X (2X Zoom). (A, C, E, G) - Untreated. (B, D, F, H) - Cells cultured overnight with 50 μM H2O2. (A, B) DAPI (blue) nuclear stain. (C, D) Phalloidin Alexa Fluor 633 F-Actin stain. (E, F) 4-Hydroxy-2-hexenal Antibody. (G, H) Composite.

Western Blot analysis of 4-hydroxy-2-hexanal-BSA Conjugate showing detection of 67 kDa 4-hydroxy-2-hexenal protein using Mouse Anti-4-hydroxy-2-hexenal Monoclonal Antibody, Clone 6F10. Lane 1: Molecular Weight Ladder (MW). Lane 2: BSA (0.5 μg). Lane 3: 4-hydroxyl nonenal-BSA (0.5 μg). Lane 4: 4-hydroxy nonenal-BSA (2.0 μg). Lane 5: 4-hydroxy-2-hexenal (0.5 μg). Lane 6: 4-hydroxy-2-hexenal (2.0 μg). Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-4-hydroxy-2-hexenal Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:2000 for 60 min at RT. Color Development: ECL solution for 5 min in RT. Predicted/Observed Size: 67 kDa.

Flow Cytometry analysis using Mouse Anti-4-hydroxy-2-hexenal Monoclonal Antibody, Clone 6F10. Tissue: Neuroblastoma cells (SH-SY5Y). Species: Human. Fixation: 90% Methanol. Primary Antibody: Mouse Anti-4-hydroxy-2-hexenal Monoclonal Antibody at 1:50 for 30 min on ice. Secondary Antibody: Goat Anti-Mouse: PE at 1:100 for 20 min at RT. Isotype Control: Non Specific IgG. Cells were subject to oxidative stress by treating with 250 μM H2O2 for 24 hours.