Rabbit anti-Goat IgG (H&L), HRP conjugated
AS09 605 | clonality: polyclonal | host: rabbit | reactivity: goat IgG (H&L)
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|Recommended dilution||1: 10 000 -1: 50 000 (ELISA), 1: 10 000 - 50 000 with regular ECL (WB), 1: 500-1: 5000 (IHC)|
|Expected | apparent MW|
|Confirmed reactivity||goat IgG heavy and light chains (H&L)|
|Predicted reactivity||goat IgG heavy and light chains (H&L)|
|Not reactive in||no confirmed exceptions from predicted reactivity known in the moment|
|Additional information||No reactivity is observed to non-immunoglobulin goat serum proteins based in immunoelectrophoresis.
BSA and milk have to be replaced by other blocking reagents, like doneky serum or commercial formulations which are free from bovine IgG.
|5 µg of total extract from Arabidopsis thaliana leaf (S) 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-BiP antibody (AS09 615) 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 (goat anti-rabbit 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 ECL Advance 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.|
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