Various lysates were subjected to SDS PAGE followed by western blot with 66200-1-Ig (Acetyl-Tubulin (Lys40) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.
(left) Confocal image of a single-plane cross-section of a healthy human islet, showing primary cilia on both alpha cells (glucagon, red) and non-alpha cells (non-red). Acetylated alpha tubulin, cilia (green, Cat. No 66200-1-Ig, 1:400), nuclei (blue), scale 20 μm. (right) Primary cilia in wildtype B6 mouse islets: acetylated alpha tubulin (green, Cat. No 66200-1-Ig, 1:400), glucagon (red), nuclei (blue), single-plane images, scales 10 μm. (Fig from Dr. Jing Hughes)
Immunofluorescent images of MDCK cells stained for IFT88 rabbit pAb (13967-1-AP) and acetylated tubulin mouse mAb (66200-1-Ig) at dilution of 1:50, further stained with Alexa Fluor 594-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) for 13967-1-AP, and Alexa Fluor 488-conjugated AffiniPure Goat anti-Mouse IgG (H+L) for 66200-1-Ig.
Immunohistochemical analysis of paraffin-embedded mouse ovary tissue slide using 66200-1-Ig (Acetyl-Tubulin (Lys40) antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
Specificity validation by indirect ELISA. Indirect ELISA was carried out by coating E.coli expressed GST-tagged Tubulin alpha (70 ng/well), non-acetylated Tubulin alpha, and Lys40 acetylated Tubulin alpha (with excessive amount) followed by blocking and adding serial diluted GST tag primary antibody (Cat.NO. 66001-2-Ig) and Acetylated Tubuliln antibody 66200-1-Ig respectively. HRP-Goat anti-mouse secondary antibody was used for detection. Signal was developed with TMB and stopped by H2SO4. Signal strength was measured by absorbance at 450 nm.
