Synthetic peptide corresponding to Mouse 15 Lipoxygenase 1 aa 50-150 conjugated to Keyhole Limpet Haemocyanin (KLH). Database link: P39654 (Peptide available as ab139428)
WB: Raw264.7 and EL4 whole cell lysates and mouse kidney, heart, small intestine and brain tissue lysates.
ICC/IF: RAW246.7 cells.
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ICC/IF image of ab87353 stained RAW246.7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab87353, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Anti-15 Lipoxygenase 1 antibody (ab87353)
All lanes : Anti-15 Lipoxygenase 1 antibody (ab87353) at 1 µg/ml
Lane 1 : Kidney (Mouse) Tissue Lysate Lane 2 : Heart (Mouse) Tissue Lysate Lane 3 : Small Intestine (Mouse) Tissue Lysate Lane 4 : RAW 264.7 (Mouse leukaemic monocyte macrophage cell line) Whole Cell Lysate Lane 5 : Brain (Mouse) Tissue Lysate Lane 6 : Forebrain (Mouse) Tissue Lysate Lane 7 : EL4 whole cell lysate (ab7183)
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 75 kDa Observed band size : 75 kDa Additional bands at : 41 kDa,74 kDa. We are unsure as to the identity of these extra bands.
Suzuki H et al. Arachidonate 12/15-lipoxygenase-induced inflammation and oxidative stress are involved in the development of diabetic cardiomyopathy. Diabetes64:618-30 (2015).
Read more (PubMed: 25187369) »