Superclonal Recombinant Secondary Antibodies

Superclonal secondary antibodies are designed to provide lot-to-lot consistency. Endogenous TOMM20 in HeLa cells were labeled with rabbit TOMM20 monoclonal primary antibody, which was then detected with four different lots of Goat anti-Rabbit IgG (Heavy Chain), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 488 (Cat. No. A55053) (green). Comparable signal and noise fluorescence intensities from these four lots demonstrate lot-to-lot consistency of Superclonal secondary antibodies. Inset represents control cells with no primary (NP) antibody to assess noise.

Superclonal secondary antibodies are designed to provide enhanced specificity. Endogenous HDAC2 in HeLa cells were labeled with mouse HDAC2 monoclonal primary antibody, which was then detected with Goat anti-Mouse IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 647 (Cat. No. A55060) (red). Simultaneously, LRP130 was labeled with rabbit LRP130 antibody followed by anti-rabbit secondary antibody conjugated to Alexa Fluor Plus 488 (green). Goat anti-Mouse IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 647 (Cat. No. A55060) specifically detects mouse primary antibody and does not cross-react with rabbit primary antibody.

Immunohistochemical analysis of Goat-anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 (Cat. No. A66724). Analysis was performed on rat testis FFPE sections. De-paraffinized sections were subjected to heat induced epitope retrieval using eBioscience IHC Antigen Retrieval Solution - Low pH (10X) (Cat. No. 00-4955-58) and blocked with 2% normal goat serum for 1 hour at room temperature. Sections were then probed for HSPA-4 overnight at 4oC. This was detected by incubating with Goat-anti-Human IgG (H+L), Superclonal Recombinant Secondary, Antibody Alexa Fluor 488 (Cat. No. A66724, used at 2 µg/mL) in 0.1% normal goat serum for 1 hour at room temperature. The images were captured on EVOS M7000 Imaging System (Cat. No. AMF7000) at 20X magnification. Panel a) represents the composite image of HSPA-4 (green) and the nuclear stain (blue). Panel b) represents the composite image of no primary antibody control and the nuclear stain (blue).

Immunohistochemical analysis of Goat anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 647 (Cat. No. A66725). Analysis was performed on rat testis FFPE sections. De-paraffinized sections were subjected to heat induced epitope retrieval using eBioscience IHC Antigen Retrieval Solution - Low pH (10X) (Cat. No. 00-4955-58) and blocked with 2% normal goat serum for 1 hour at room temperature. Sections were then probed for HSPA-4 overnight at 4oC. This was detected by incubating with Goat anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 647 (Cat. No. A66725, used at 2 µg/mL) in 0.1% normal goat serum for 45 minutes at room temperature. The images were captured on EVOS M7000 Imaging System (Cat. No. AMF7000) at 20X magnification. Panel a) represents the composite image of HSPA-4 (red) and the nuclear stain (blue). Panel b) represents the composite image of no primary antibody control and the nuclear stain (blue).

Immunohistochemical analysis of Goat anti-Human IgG Fc, Recombinant Secondary Antibody, Alexa Fluor Plus 488 (Cat. No. A66726). Analysis was performed on rat testis FFPE sections. De-paraffinized sections were subjected to heat induced epitope retrieval using eBioscience IHC Antigen Retrieval Solution - Low pH (10X) (Cat. No. 00-4955-58) and blocked with 2% normal goat serum for 1 hour at room temperature. Sections were then probed for HSPA-4 overnight at 4oC. This was detected by incubating with Goat anti-Human IgG Fc, Recombinant Secondary Antibody, Alexa Fluor Plus 488 (Cat. No. A66726, used at 2 µg/mL) in 0.1% normal goat serum for 45 minutes at room temperature. The images were captured on EVOS M7000 Imaging System (Cat. No. AMF7000) at 20X magnification. Panel a) shows the specific detection of HSPA-4 (green) in the nucleus and cytoplasm of Leydig cells and seminiferous duct cells in testis. Panel b) shows absence of non-specific staining in the presence of secondary antibody alone. Panel c) represents the composite image of HSPA-4 (green) and the nuclear stain (blue). Panel d) represents the composite image of no primary antibody control and the nuclear stain (blue).

Immunofluorescence analysis comparing the Alexa Fluor 488 and Alexa Fluor Plus 488 conjugates of Goat-anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody. Analysis was performed on fixed and permeabilized T-47D cells blocked with 2% BSA at room temperature and stained for MUC-1 protein using Gatipotuzumab Recombinant Human Monoclonal Antibody (Cat. No. MA5-42157, used at 1:500 dilution) in 0.1% BSA overnight at 4-degree Celsius. This was detected by incubating with Goat-anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 (Cat. No. A66724) or Goat anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 488 (Cat. No. A56021) at 1:1,000 dilution. Panel a) represents the composite images of cells that were stained for detection and localization of MUC-1 protein using Goat-anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor 488 (green) and the nuclear counterstain (red). Panel b) represents the composite images of cells that were stained for detection and localization of MUC-1 protein using Goat anti-Human IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 488 (green) and the nuclear counterstain (red). The images were captured at 20X magnification on CellInsight CX7 LZR High Content Analysis Platform (Cat. No. CX7A1110LZR).