|Storage Buffer||PBS pH 7.4, 50% glycerol, 0.09% Sodium azide *Storage buffer may change when conjugated|
|Storage Temperature||-20ºC, Conjugated antibodies should be stored at 4ºC|
|Shipping Temperature||Blue Ice or 4ºC|
|Purification||Protein G Purified|
|Cite This Product||StressMarq Biosciences Cat# SMC-504, RRID: AB_2702711|
|Certificate of Analysis||A 1:1000 dilution of SMC-504 was sufficient for detection of Acrolein in 2 µg of Acrolein conjugated to BSA by ECL immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary Antibody.|
|Alternative Names||Acrolein modified protein Antibody, Acrolein conjugated protein Antibody, 2-Propen-1-one Antibody, 2-propenal Antibody, Acraldehyde Antibody, Acrolein Antibody, Acrylic aldehyde Antibody, Protein-bound Acrolein Antibody|
|Research Areas||Alzheimer's Disease, Cancer, Lipid peroxidation, Neurodegeneration, Neuroscience, Oxidative Stress|
|Scientific Background||Lipid peroxidation occurs when oxidizing agents attack carbon-carbon double bonds found in unsaturated lipids. In addition to membrane degradation, oxidation end-products have been found to damage cell viability through their mutagenic and toxic properties. These downstream functional consequences facilitate the development of disease and premature aging. Acrolein is an electrophilic conjugated aldehyde that is a terminal product of lipid peroxidation. Acrolein is highly mutagenic and reacts with nucleophilic functional groups in DNA and proteins such as cysteine, histidine, and lysine residues (1).|
|References||1. Takabe, W. et al. (2001) Carcinogenesis. 22 (6): 935-941|
Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Acrolein Monoclonal Antibody, Clone 2H2 (SMC-504). Tissue: Embryonic kidney epithelial cell line (HEK293). Species: Human. Fixation: 5% Formaldehyde for 5 min. Primary Antibody: Mouse Anti-Acrolein Monoclonal Antibody (SMC-504) at 1:50 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) Acrolein Antibody. (G,H) Composite. Courtesy of: Dr. Robert Burke, University of Victoria.
Western Blot analysis of Acrolein-BSA Conjugate showing detection of 67 kDa Acrolein protein using Mouse Anti-Acrolein Monoclonal Antibody, Clone 2H2 (SMC-504). Lane 1: Molecular Weight Ladder (MW). Lane 2: Acrolein-BSA (0.5 µg). Lane 3: Acrolein-BSA (2.0 µg). Lane 4: BSA (0.5 µg). Lane 5: BSA (2.0 µg). Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-Acrolein Monoclonal Antibody (SMC-504) 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-Acrolein Monoclonal Antibody, Clone 2H2 (SMC-504). Tissue: Neuroblastoma cells (SH-SY5Y). Species: Human. Fixation: 90% Methanol. Primary Antibody: Mouse Anti-Acrolein Monoclonal Antibody (SMC-504) 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.
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