Anti au1 antibody

Expression of adenylyl cyclase related proteins were analyzed using
Agilent Gene Expression Microarray technique by PhalanxBio Inc. (CA, San Diego).
C1C2 protein was detected by anti-AU1 antibody (Fitzgerald, 1:2,000) or
anti-AC5/6 antibody (Santa Cruz, 1:200). Additional antibodies included: GAPDH
(Fitzgerald, 1:20,000); cMLCK (Abgent, 1:1,000); MLC2v (Synaptic Systems,
1:1,000) PKA catalytic subunit (BD Transduction, 1:1,000); p-ERK1/2, p38,
p-PKARII α and β (Santa Cruz Biotech, 1:200); PLB (Affinity
Bioreagents, 1:5,000); phospho-16-PLB (Badrilla, 1:3,000); S100A1 (Epiyomics,
1:1,000); SERCA2a antibody (Enzo, 1:1,000); phospho-αB-crystallin
(CryAB) and total CryAB antibodies (Enzo, 1:1,000); P-308-Akt, P-473-Akt, T-Akt,
p-GSK3a/b, p-MDM2, P-p70S6K, T-p70S6K, and phospho-S22/23 troponin I (TnI) (Cell
Signaling, 1:1,000 each); Vinculin (Sigma, 1:100,1000).

Isolated cardiac myocytes were attached to laminin coated 2-well chamber
slides for 1 hr, washed, fixed (10% formalin, 15 min, 23°C),
blocked with normal goat serum (1 hr) and incubated (4°C, overnight)
with: anti-AU1 antibody (Fitzgerald, 1:300; for detecting C1C2 transgene
protein) and anti-Cav3 antibody (BD Pharmagen, 1:100; for detecting caveolae).
Cardiac myocytes were washed with PBS and then incubated with secondary
antibodies (Alexia Fluo 488 or 594 conjugated, 1:1000 dilution) for 1 hr. To
identify the nucleus, cells were incubated with Hoechst dye (1:1000 dilution, 20
min). Cardiac myocytes then were imaged as previously described.^{18 (link)}