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Rabbit anti-Goat IgG (H&L), HRP conjugated

AS09 605 | Clonality: Polyclonal  |  Host: Rabbit  | Reactivity: Goat IgG (H&L)
Rabbit anti-Goat IgG (H&L), HRP conjugated in the group Secondary Antibodies / Anti-Goat / HRP (horse radish peroxidase) at Agrisera AB (Antibodies for research) (AS09 605)

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Product Information

Immunogen Purified goat IgG, whole molecule
Host Rabbit
Clonality Polyclonal
Purity Affinity purified using solid phase goat IgG
Format Lyophilized
Quantity 1 mg
Reconstitution For reconstitution add 1.1 ml of sterile water. Let it stand 30 minutes at room temperature to dissolve. Centrifuge to remove any particulates. Prepare fresh working dilutions daily.
Storage Store lyophilized material at 2-8°C.
For long time storage after reconstitution, dilute the antibody solution with glycerol to a final concentration of 50% glycerol and store as liquid at -20°C, to prevent loss of enzymatic activity.
For example, if you have reconstituted 1 mg of antibody in 1.1 ml of sterile water add 1.1 ml of glycerol. Such solution will not freeze in -20°C. If you are using a 1:5000 dilution prior to diluting with glycerol, then you would need to use a 1:2500 dilution after adding glycerol. Prepare working dilution prior to use and then discard. Be sure to mix well but without foaming.
Tested applications ELISA (ELISA), Immunohistochemistry (IHC), Western blot (WB)
Recommended dilution 1 : 10 000 -1 : 50 000 (ELISA), 1 : 500-1 : 5000 (IHC), 1 : 10 000 - 50 000 (WB)

Reactivity

Confirmed reactivity Goat IgG heavy and light chains on all goat immunoglobulins
Not reactive in Non-immunoglobulin goat serum proteins based in immunoelectrophoresis.

Application examples

Application examples Application example

western blot using secondary antibody rabbit anti-goat immunoglobulins, HRP conjugated

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, AS09 602) diluted to 1:50 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent in extreme low femtogram range, 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.

Additional information

Additional information HRP-conjugate is supplied in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 % (w/v) BSA, Protease/IgG free with 0.1 % (v/v) of ProClin 150 as preservative.
For blocking BSA and non-fat milk is recommended to be replaced by other blocking reagents, like donkey serum or commercial formulations which are free from bovine IgG.

Related products

Related products AS09 612 | Rabbit anti-goat IgG (H&L) biotin conjugated, secondary antibodies
AS09 630 | Rabbit anti-goat IgG (H&L), ALP conjugated, secondary antibodies

Collection of secondary antibodies

Background

Background Rabbit anti-goat IgG is a secondary antibody conjugated to HRP (horse radish peroxidase) which binds to all goat immunoglobulins in immunological assays.

Product citations

Selected references Sinclair et al. (2017) Etiolated Seedling Development Requires Repression of Photomorphogenesis by a Small Cell-Wall-Derived Dark Signal. Curr Biol. 2017 Nov 20;27(22):3403-3418.e7. doi: 10.1016/j.cub.2017.09.063.

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