This antibody does not cross-react with Acrolein, Crotonaldehyde, 4-Hydroxy-2-hexenal, 4-Hydroxy nonenal, Malondialdehyde, or Methylglyoxal modified proteins.
Immunohistochemistry analysis of rat heart, fixed in formalin. The Primary Antibody used was Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725) at 1:25 for 1 hour at room temperature. The secondary antibody used was Goat Anti-Mouse IgG: Alexa Fluor 488. Counterstain: DAPI (blue) nuclear stain. Magnification: 63X.(A) DAPI (blue) nuclear stain. (B) Actin (C) Hexanoyl-Lysine adduct Antibody (D) Composite.
Flow cytometry analysis of human neuroblastoma cells (SH-SY5Y), fixed in 90% methanol, using Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725) at 1:50 for 30 minutes on ice. The secondary antibody used was Goat Anti-Mouse: PE at 1:100 for 20 minutes at room temperature. Isotype Control: Non Specific IgG. Cells were subject to oxidative stress by treating with 250 µM H2O2 for 24 hours.
Western Blot - Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725)
Western blot analysis of human Cervical cancer cell line (HeLa) lysate showing detection of Hexanoyl-Lysine adduct protein using Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725) at 1:1,000 for 2 hours at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: HeLa cell lysate. Lane 3: H2O2 treated HeLa cell lysate. Load: 12 µg. Block: 5% Skim Milk in TBST. The secondary antibody used was Goat Anti-Mouse IgG: HRP at 1:2000 for 60 minutes at room temperature. Color Development: ECL solution for 5 minutes in room temperature.
Western Blot - Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725)
Western blot analysis of Hexanoyl Lysine-BSA Conjugate showing detection of 67 kDa Hexanoyl-Lysine adduct protein using Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725) at 1:1,000 for 2 hours at room temperature. Lane 1: Molecular Weight Ladder (MW). Lane 2: Hexanoyl Lysine-BSA. Lane 3: BSA. Load: 0.5 µg. Block: 5% Skim Milk in TBST. The secondary antibody used was Goat Anti-Mouse IgG: HRP at 1:2000 for 60 minutes at room temperature. Color Development: ECL solution for 5 minutes in room temperature. Predicted/Observed Size: 67 kDa.
Immunocytochemistry/Immunofluorescence analysis of human embryonic kidney epithelial cell line (HEK293), fixed in 5% formaldehyde for 5 minutes, using Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725), at 1:50 for 30-60 min at room temperature. The secondary antibody used was Goat Anti-Mouse Alexa Fluor 488 at 1:1500 for 30-60 min at room temperature. Counterstain: Phalloidin Alexa Fluor 633 F-Actin stain; DAPI (blue) nuclear stain at 1:250, 1:50,000 for 30-60 min at room temperature. 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) Hexanoyl-Lysine adduct Antibody. (G,H) Composite.
Immunohistochemistry analysis of rat kidney. The Primary Antibody used was Anti-Hexanoyl-Lysine adduct Antibody [5D9] (A304725) at 1:100 for Overnight at 4°C, then 30 minutes at 37°C. The secondary antibody used was Goat Anti-Mouse IgG (H+L): FITC for 45 minutes at 37°C. Counterstain: DAPI for 3 min at room temperature. Magnification: 10X.
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