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

PRODUCT INFORMATION IN PDF

product information background   Goat anti-rabbit IgG is a secondary antibody conjugated to HRP which binds to all rabbit IgGs in immunological assays. immunogen   purified rabbit IgG, whole molecule antibody format   goat polyclonal affinity purified goat IgG Lyophilized quantity   1 mg storage   Store lyophilized material at 2-8°C. For storage at -20°C after reconstitution dilute antibody solution with an equal volume of glycerol to obtain final glycerol concentration of 50 % 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. related products   AS09 607 | Goat anti-rabbit IgG (H&L) AP conjugated AS09 608 | Goat anti-rabbit IgG (H&L) biotin conjugated additional information   Antibody concentration is 1 mg/ml and it has been purified by antigen-specific chromatography HRP-conjugate is supplied in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 % (w/v) BSA, Protease/IgG free 0.1 % (v/v) of Kathon CG is used as preservative.

application information
recommended dilution		1: 50 000 -1: 90 000 (ELISA), 1 : 75 000 with enhanced ECL and 1: 25 000 with regular ECL (WB), 1: 500 -1: 5000 (IHC)
selected references		to be added when available

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-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, 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.

Comparison of Agrisera 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 standard ECL 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 enhanced ECL and 1: 25 000 with regular ECL (WB), 1: 500 -1: 5000 (IHC).

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