Goat anti-Chicken IgY (H&L), HRP conjugated

Application example

5 µg of total extract from (1) Hordeum vulgare total leaf,  (2) Zea mays  (3) Spinacia oleracea extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary anti-PsbA antibody (AS01 016) at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-hen IgG horse radish peroxidase conjugated, AGRISERA) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescen detection reagent according to the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds. 

5 µg of total protein from  A.thaliana (1), H. vulgare (2),  Z.mays (3),  S. oleracea (4), extracted with Agrisera PEB extraction buffer (AS08 300) were separated on  4-12% SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in  for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-hen IgY horse radish peroxidase conjugated, from  Agrisera AS09 603) diluted to 1:50 000  for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL  detection reagent according to the manufacturers instructions.  Exposure time was 5 seconds.

Concentration: 1.0 mg/ml

This HRP-conjugate is supplied in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free 0.1 % (v/v) of Kathon CG is used as preservative.

Antibody binds to:
heavy chains on chicken IgY
light chains on all chicken immunoglobulins

No reactivity is observed to non-immunoglobulin chicken serum proteins based in immunoelectrophoresis.

Chicken immunoglobulin is often called hen or chicken IgG, however it is derived from egg yolk, therefore IgY.

Goat anti-chicken IgY (H&L) is a secondary antibody conjugated to HRP (horse radish peroxidase) which binds to chicken IgY in immunological assays.