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

AS09 602 | Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)

Product Information

Immunogen

Purified Rabbit IgG, whole molecule.

Host Goat

Clonality Polyclonal

Purity Affinity purified 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. 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 : 50 000 -1 : 90 000 (ELISA), 1 : 500 -1 : 5000 (IHC), 1: 10 000 -1 : 20 000 (WB)

Reactivity

Confirmed reactivity Based on IEP, this antibody Reacts with: Rabbit IgG heavy chainslight chains on all Rabbit immunoglobulins

Application examples

Application examples Application example

western blot detection of PsaC using Agrisera primary and secondary antibodies

5 µg of total extract from (1) Hordeum vulgaretotal 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-PsaC antibody (AS04 042) 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, AS09 602, 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 chemiluminescent 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.

Comparison of Agrisera secondary antibody sensitivity

comparison of secondary antibody sensitivity

10 μg of mitochondrial fraction from Arabidopsis thaliana (1,3) and Arabidopsis thaliana leaf extract (2,4) were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 1000 anti-COXII antibodies (2h in TBST) followed by incubation with 1: 10 000 secondary anti-rabbit (1h) HRP-coupled antibodies from Agrisera (left panel) and other manufacture (right panel) and visualized with chemiluminescent detection reagent, on Kodak autoradiography film for 5 s. Antibody in left panel detects target protein also in total cell extract (2) and can be used in higher dilution than applied 1: 10 000.

Agrisera goat anti-rabbit HRP conjugated antibody (AS09 602) can be used at following dilutions: 1: 50 000 -1: 90 000 (ELISA), 1 : 75 000 with chemiluminescence detection range of extreme low picogram and 1: 25 000 with chemiluminescence detection reagent of mid femtogram (WB), 1: 500 -1: 5000 (IHC).

Additional information

Antibody is provided in: 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1% BSA (w/v), Protease IgG free, 0.1 % (v/v) Kathon CG.

Affinity purified antibody is > 95 % pure, according to SDS-PAGE.

This antibody is used on a very wide range of samples from various species including many model plants, algae and diatoms.

No reactivity is observed to non-immunoglobulin rabbit serum.

Related products

AS09 607 | Goat anti-rabbit IgG (H&L) ALP conjugated secondary antibodies
AS09 608 | Goat anti-rabbit IgG (H&L) biotin conjugated secondary antibodies
AS10 668 | Goat anti-Rabbit IgG (H&L) HRP conjugated, min.cross-reactivity to bovine/Human/mouse IgG/serum secondary antibodies

Background

Goat anti-rabbit IgG (H&L) is a secondary antibody conjugated to HRP which binds to all rabbit IgGs in immunological assays.

Product citations

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Tyuereva et al. (2017). The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants. Photosynth Res. 2017 Apr 5. doi: 10.1007/s11120-017-0376-9. (Hordeum vulgare, western blot)
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CiteAB has listed this antibody as one of the 7000 most published antibodies in the world.