Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography. They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent. The average molecular weight is reported to be about 160 kDa. The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective.
Horseradish peroxidase (HRP) conjugates are prepared by a modified Nakane and Kawaoi procedure (J. Histochem. Cytochem. 1974. 22, 1084). Peroxidase conjugates are commonly used for immunohistochemistry, Western blotting, and ELISA. Affinity-purified anti-horseradish peroxidase and conjugates are available for detection of horseradish peroxidase antigen or for signal amplification of HRP-containing reagents. For immunostaining of mammalian cells, an advantage of using anti-horseradish peroxidase is reduced background, since the antibody does not recognize the endogenous peroxidase-like enzymes found in those cells.
Goat anti-Rabbit IgG-HRP antibody
WB, ELISA, IHC
Rabbit IgG (H+L).
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
1*TBS (pH7.4), 0.5%BSA, 50%Glycerol.
Protein affinity purified.
Fig1: Western blot analysis of rabbit anti-GAPDH antibody (ET1601-4,1/2000) and rabbit anti-GST3 antibody (ER1802-63, 1/1000) with K562 Cell and A549 Cell lysates (All lanes proteins at 10 µg per lane). Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody was used in 5% BSA at room temperature for 2 hours. Goat anti-rabbit IgG -HRP) (HA1001) was used for 1 hour at room temperature at 1:40,000 (Lane A), 1:80,000 (Lane B), 1:160,000 (Lane C) and 1:320,000 (Lane D).
Exposure time: 60 seconds
Fig2: The Cross-reactions of the secondary antibody (HA1001) with Mouse IgG and Human IgG.
Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. Goat anti-rabbit IgG -HRP (HA1001) was used for 1 hour at room temperature at 1:40,000 (Lane A) and 1:180,000 (Lane B).
Lane A: Mouse IgG (30 ng per lane).
Lane B: Human IgG (30 ng per lane).
Exposure time: 2 minutes
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