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Animal cell antibodies / Secondary antibodies/blocking / Anti-rabbit / HRP (horseradish peroxidase)


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

Art no: AS09 602
Price: 150
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product information

background  

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

antibody format  

goat

polyclonal

affinity purified goat IgG

lyophilized

quantity  

1 mg

storage  

Store at 2-8°C. For extended storage after rehydration, add an equal volume of glycerol and store at -20°C. Shelf life is one year from a date of receipt.

related products  

AS09 607 | Goat anti-rabbit IgG (H&L) AP conjugated

AS09 608 | Goat anti-rabbit IgG (H&L) biotin conjugated

recommended dilution

1: 50 000 -1: 90 000 (ELISA), 1 : 75 000 with ECL Advance and 1: 25 000 with regular ECL (WB), 1: 500 -1: 5000 (IHC)

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.

No reactivity is observed to non-immunoglobulin rabbit serum.


application example

 

western blot detection of PsaC using Agrisera primary and secondary antibodies

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

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 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.



||| For applications or usage on species others than stated above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at  support@agrisera.com



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