Secondary Antibody
Secondary Antibody Definition
The binding of antibodies to target antigens can be visualized by either colorimetric or fluorescent methods. The methods for visualization involve either direct or indirect binding. Primary antibodies can be directly conjugated to fluorophores, enzymes or other substances that will aid in the visualization, such as GFP, biotin or horseradish peroxidase. However, when primary antibodies are directly conjugated, this limits the number of applications that they will be suitable for, as well as the sensitivity. For this reason, most researchers prefer to use conjugated secondary antibodies that will bind to the primary antibody of interest.
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How to choose Secondary Antibodies
Secondary antibodies are raised against the same host species that was used to generate the primary antibody. For example, if the primary antibody was raised in rabbit, then an anti-rabbit secondary antibody that was raised in a species other than rabbit should be used. Secondary antibodies that are designed for use in Western blots and ELISA are generally conjugated to enzymes, such horseradish peroxidase or alkaline phosphatase. Biotin-conjugated secondary antibodies are also commonly used. The biotin label is small and stable, so it rarely interferes with the function of the labeled antibody. Biotin binds very strongly to avidin, and the biotin-avidin interaction can be used to further amplify the signal.
Streptavidin is purified from the bacterium Streptomyces avidinii, and although this protein does not bind quite as strongly to biotin, it is used in various detection systems because it is unglycosylated and less likely to aggregate. Streptavidin also exhibits lower non-specific binding than avidin. Each avidin or streptavidin will bind four biotins per molecule. When a biotinylated secondary antibody is used for detection, the signal can be significantly amplified by incubation with an avidin-biotin complex or a labeled streptavidin-biotin. Streptavidin can also be conjugated to detection enzymes, such as horseradish peroxidase or alkaline phosphatase, as well as fluorophores, such as GFP and R-Phycoerythrin.
While secondary antibodies can be valuable tools for amplifying signals in immunohistochemistry, additional steps are often required to prevent non-specific binding, which can make it difficult to visualize the exact location of the protein of interest. The causes of non-specific binding include interactions between the primary and/or secondary antibody and serum proteins, ionic interactions between antibodies and tissues, as well as interactions with endogenous molecules that may interfere with the detection system used.
Non-specific hydrophobic interactions can be at least partially prevented by incubating tissue sections with heat-inactivated normal serum or bovine serum albumin. However, it is important to choose a normal serum that will not interact with the primary or secondary antibodies. The best choice is generally a serum from the same host animal as the secondary antibody or a serum from a species that is unrelated to either host species. Ionic interactions can be reduced by increasing the ionic strength of the fixative and diluents. Endogenous enzyme interference can be reduced through the use of blocking reagents, such as hydrogen peroxide (for horseradish peroxidase) and levamisole (for alkaline phosphatase). When avidin-biotin based detection systems are used, interference from endogenous biotin can be reduced by first incubating the sample with avidin to block the biotin binding sites and then incubating the sample with biotin to block any additional biotin binding sites on the avidin molecule.
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anti-Mouse IgG+HRP
Host/Reactivities:
Host: Goat / Reactivity: Mouse
Applications:
Western Blot (WB)
Goat anti Rabbit IgG, Fc Fragment Specific Antibody, Peroxidase Conjugated
Host/Reactivities:
Host: Goat
Purification:
Immunoaffinity chromatography
Applications:
ELISA, Western Blot, Immunohistochemistry, Immunocytochemistry
Goat Anti-Mouse IgG Heavy and Light Chain Antibody, Peroxidase Conjugated
Host/Reactivities:
Host: Goat
Purification:
Immunoaffinity chromatography
Applications:
ELISA, Western Blot, Immunohistochemistry, Immunocytochemistry
Goat Anti-Mouse IgG Heavy and Light Chain Antibody, Alkaline Phosphatase Conjugated
Host/Reactivities:
Host: Goat
Purification:
Immunoaffinity chromatography
Applications:
ELISA, Western Blot, Immunohistochemistry, Immunocytochemistry
Biotin Conjugated Goat Anti-human IgM secondary antibody
Host/Reactivities:
Host: Goat
Isotype/Clone:
Isotype: IgM
Applications:
Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB), Direct ELISA (EIA)
Rabbit anti-Human IgG Fc fragment polyclonal Antibody, HRP conjugated
Other Names:
fragment crystallizable region
Host/Reactivities:
Host: Rabbit / Reactivity: Human
Purification:
Caprylic Acid Ammonium Sulfate Precipitation purified
Isotype/Clone:
Isotype: IgG
Applications:
ELISA (EIA), Western Blot (WB)
Mouse secondary antibody to human IgG (Fc)-HRP
Other Names:
Mouse anti-human IgG (Fc)-HRP conjugated; Mouse anti-human IgG
Host/Reactivities:
Host: Mouse / Reactivity: Human
Purification:
This product was purified from antiserum by immunoaffinity chromatography (monoclonal).
Isotype/Clone:
Isotype: IgG
Applications:
Western Blot (WB), ELISA (EIA)
Donkey anti-Rabbit IgG (H&L) - Affinity Pure, DyLight350 Conjugate, min x w/bovine, chicken, goat, guinea pig, hamster, horse, human,mouse, rat or sheep IgG
Other Names:
Donkey anti-Rabbit IgG (H&L); Affinity Pure
Host/Reactivities:
Host: Donkey / Reactivity: Rabbit.Cross Reactivity: Based on IEP, no reactivity is observed to:non-immunoglobulin rabbi...
Purification:
Affinity purified antibody is > 95% based on SDS-PAGEAffinity purified using solid phase Rabbit IgG.
Applications:
Flow Cytometry (FC/FACS), Immunofluorescence (IF), ELISA (EIA), Immunomicroscopy (IM)
Sheep anti Human IgG3 (subclass specific)
Host/Reactivities:
Host: Sheep / Reactivity: Human
Applications:
Precipitation assays
Mouse secondary antibody to human IgG (Fc)-Unconjugated
Other Names:
Mouse anti-human IgG (Fc)-Unconjugated; Mouse anti-human IgG
Host/Reactivities:
Host: Mouse / Reactivity: Human
Purification:
This product was purified from antiserum by immunoaffinity chromatography (monoclonal).
Isotype/Clone:
Isotype: IgG
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), Immunofluorescence (IF), ELISA (EIA)
Goat anti Human IgM (mu chain) (Alk Phos)
Other Names:
Goat anti Human Immunoglobulin M (mu chain) secondary (Alk Phos)
Host/Reactivities:
Host: Goat / Reactivity: No reactivity is observed to non-immunoglobulin Human serum proteins or light chains on all Human im...
Purification:
Affinity purified using solid phase Human IgM
Applications:
User optimized
HRP Rabbit Anti-Camel IgG (H+L) Antibody
Other Names:
HRP Rabbit Anti-Camel IgG (H+L): N/A
Host/Reactivities:
Host: Rabbit
Purification:
Affinity Purification
Isotype/Clone:
Isotype: Horsedaish peroxidase conjugated IgG
Applications:
Western Blot (WB)
Biotin Conjugated Goat Anti-rabbit IgG secondary antibody
Host/Reactivities:
Host: Goat
Isotype/Clone:
Isotype: IgG
Applications:
Immunohistochemistry (IHC), Immunocytochemistry (ICC), Western Blot (WB), ELISA (EIA)
Rabbit anti-Sheep IgG polyclonal Antibody, HRP conjugated
Host/Reactivities:
Host: Rabbit / Reactivity: Sheep
Purification:
Caprylic Acid Ammonium Sulfate Precipitation Purified
Isotype/Clone:
Isotype: IgG
Goat anti Rabbit IgG
Other Names:
Goat anti Rabbit Immunoglobulin G secondary
Host/Reactivities:
Host: Goat / Reactivity: Rabbit
Purification:
Goat anti Rabbit IgG was purified by affinity chromatography.
Applications:
User optimized
Goat Anti-Rabbit IgM-HRP
Other Names:
Goat Anti-Rabbit IgM - HRP; Goat Anti-Rabbit IgM (u heavy chain specific)
Host/Reactivities:
Host: Goat / Reactivity: Rabbit
Purification:
Affinity chromatography on rabbit IgM covalently linked to agarose
Isotype/Clone:
Isotype: IgG
Applications:
ELISA; Immunoblot; Immunofluorescence; Immunohistochemistry
Rabbit anti-Mouse IgG Fc fragment polyclonal Antibody, HRP conjugated
Host/Reactivities:
Host: Rabbit / Reactivity: Mouse
Purification:
Caprylic Acid Ammonium Sulfate Precipitation Purified
Isotype/Clone:
Isotype: IgG
Mouse Anti-Human IgG (Fc)-HRP
Other Names:
Mouse Anti-Human IgG (Fc) - HRP; Mouse Anti-Human IgG (Fc) (gamma chain specific)
Isotype/Clone:
Isotype: Mouse IgG2bK / Clone H2
Mouse secondary antibody to human IgG (Fc)-Biotin
Other Names:
Mouse anti-human IgG (Fc)-Biotin conjugated; Mouse anti-human IgG
Host/Reactivities:
Host: Mouse / Reactivity: Human
Purification:
This product was purified from antiserum by immunoaffinity chromatography (monoclonal).
Isotype/Clone:
Isotype: IgG
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), ELISA (EIA)
anti-Goat IgG+HRP
Host/Reactivities:
Host: Rabbit / Reactivity: Goat
Applications:
Western Blot (WB)
Anti-Human IgM Monoclonal Antibody (PE/Cyanine5 Conjugated) [MHM-88]
Other Names:
Human IgM; Immunoglobulin heavy constant mu; IGHM; Immunoglobulin M; Ig mu chain C region
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: IgG1, kappa / Clone MHM-88
Applications:
Flow Cytometry (FC/FACS)
HRP Goat Anti-Mouse IgG (H+L) Antibody
Other Names:
HRP Goat Anti-Mouse IgG (H+L): N/A
Host/Reactivities:
Host: Goat
Purification:
Affinity Purification
Isotype/Clone:
Isotype: Horseradish peroxidase conjugated IgG
Applications:
Western Blot (WB), ELISA (EIA)
HRP Goat Anti-Rabbit IgG (H+L) Antibody
Other Names:
HRP Goat Anti-Rabbit IgG (H+L): N/A
Host/Reactivities:
Host: Goat / Reactivity: Rabbit
Purification:
Affinity Purification
Isotype/Clone:
Isotype: Horseradish peroxidase conjugated IgG
Applications:
ELISA, Western Blot (WB), Immunohistochemistry (IHC)
Rabbit anti Human IgG (Alk Phos)
Other Names:
Rabbit anti Human Immunoglobulin G secondary (Alk Phos)
Host/Reactivities:
Host: Rabbit / Reactivity: Human IgG
Purification:
Rabbit anti Human IgG (Alk Phos) was purified by affinity chromatography.
Applications:
ELISA (EIA), Immunohistochemistry (IHC), Western Blot (WB)
Mouse Anti-Human IgG2 (Fc)-HRP
Other Names:
Mouse Anti-Human IgG2 (Fc) - HRP; Mouse Anti-Human IgG2 (gamma 2 chain specific)
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: Mouse (BALB/c) IgG1 kappa / Clone 31-7-4
Applications:
ELISA; FLISA
Mouse Anti-Human IgG4 (Fc)-BIOT
Other Names:
Mouse Anti-Human IgG4 (Fc) - Biotin; Mouse Anti-Human IgG4 (gamma 4 chain specific)
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: Mouse (BALB/c) IgG1 kappa / Clone HP6025
Anti-Human IgM Monoclonal Antibody (PE/Cyanine7 Conjugated) [MHM-88]
Other Names:
Human IgM; Immunoglobulin heavy constant mu; IGHM; Immunoglobulin M; Ig mu chain C region
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: Mouse IgG1, κ / Clone MHM-88
Applications:
Flow Cytometry (FC/FACS)
AP-Goat Anti-Rabbit IgG (H+L) Antibody
Other Names:
AP Goat Anti-Rabbit IgG (H+L) : N/A
Host/Reactivities:
Host: Goat
Purification:
Affinity Purification
Applications:
Western Blot (WB)
HRP Goat Anti-Rat IgG (H+L) Antibody
Other Names:
HRP Goat Anti-Rat IgG (H+L): N/A
Host/Reactivities:
Host: Goat
Purification:
Affinity Purification
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC)
FITC Goat Anti-Rabbit IgG (H+L) Antibody
Other Names:
FITC Goat Anti-Rabbit IgG (H+L): N/A
Host/Reactivities:
Host: Goat
Purification:
Affinity Purification
Isotype/Clone:
Isotype: Fluorescein conjugated IgG
Applications:
Immunofluorescence (IF), Flow Cytometry (FC) , Immunocytochemistry (ICC)
FITC Goat Anti-Rat IgG (H+L) Antibody
Other Names:
FITC Goat Anti-Rat IgG (H+L): N/A
Host/Reactivities:
Host: Goat
Purification:
Affinity Purification
Applications:
Immunocytochemistry (ICC), Flow Cytometry (FC/FACS)
Rabbit anti-Mouse IgG Fc fragment polyclonal Antibody, FITC conjugated
Host/Reactivities:
Host: Rabbit / Reactivity: Mouse
Purification:
Caprylic Acid Ammonium Sulfate Precipitation Purified
Isotype/Clone:
Isotype: IgG
Anti-Human IgD Monoclonal Antibody (FITC Conjugated)
Other Names:
Immunoglobulin heavy constant delta; IGHD; Ig delta chain C region
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: IgG2a, kappa / Clone IA6-2
Applications:
Flow Cytometry (FC/FACS)
Anti-Human IgD Monoclonal Antibody (AF488 Conjugated)
Other Names:
Human IgD; Immunoglobulin heavy constant delta; IGHD; Ig delta chain C region
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: IgG2a, kappa / Clone IA6-2
Applications:
Flow Cytometry (FC/FACS)
Mouse Anti-Human IgG2 (Fc)-PE
Other Names:
Mouse Anti-Human IgG2 (Fc) - Phycoerythrin-Labeled; Mouse Anti-Human IgG2 (gamma 2 chain specific)
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: Mouse (BALB/c) IgG1 Kappa / Clone HP6002
HRP Donkey Anti-Mouse IgG (H+L) Antibody
Other Names:
HRP Donkey Anti-Mouse IgG (H+L): N/A
Host/Reactivities:
Host: Mouse
Purification:
Affinity Purification
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC)
Goat anti Rabbit IgG (Alk Phos)
Other Names:
Goat anti Rabbit Immunoglobulin G secondary (Alk Phos)
Host/Reactivities:
Host: Goat / Reactivity: Rabbit IgG
Purification:
Goat anti Rabbit IgG (Alk Phos) was purified by affinity chromatography.
Applications:
ELISA (EIA), Immunohistochemistry (IHC), Western Blot (WB)
Goat anti-Llama IgG (H&L) - Affinity Pure, Biotin Conjugate
Other Names:
Goat anti-Llama IgG (H&L) - Affinity Pure; immunoglobulin heavy chain variable region; partial
Host/Reactivities:
Host: Goat / Reactivity: Llama.Cross Reactivity: Based on IEP, no reactivity is observed to non-immunoglobulin llama seru...
Purification:
Affinity purified antibody is > 95% based on SDS-PAGEAffinity purified using solid phase Llama IgG.
Applications:
ELISA (EIA), Immunohistochemistry (IHC)
Rabbit anti-Mouse IgG Fc fragment polyclonal Antibody, Biotin conjugated
Host/Reactivities:
Host: Rabbit / Reactivity: Mouse
Purification:
Caprylic Acid Ammonium Sulfate Precipitation Purified
Isotype/Clone:
Isotype: IgG
HRP Donkey Anti- Rabbit IgG (H+L) Antibody
Other Names:
HRP Donkey Anti-Rabbit IgG (H+L): N/A
Host/Reactivities:
Host: Donkey
Purification:
Affinity Purification
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC)
HRP, Mouse Anti-Rabbit IgG HCS
Other Names:
Mouse Anti-Rabbit IgG
Host/Reactivities:
Host: Mouse / Reactivity: Rabbit
Purification:
Affinity purified using solid phase Rabbit IgG(H+L) light chain with finally >95% purity based on SDS-PAGE
Isotype/Clone:
Isotype: Mouse IgG
Applications:
Western Blot (WB), Immunoprecipitation (IP)
Rhodamine (TRITC) -Goat Anti-Rabbit IgG (H+L) Antibody
Other Names:
Rhodamine (TRITC) -Goat Anti-Rabbit IgG (H+L): N/A
Host/Reactivities:
Host: Goat / Reactivity: Rabbit
Purification:
Affinity Purification
Isotype/Clone:
Isotype: TRITC conjugated IgG
Applications:
Immunohistochemistry (IHC), Immunocytochemistry (ICC), Flow Cytometry (FC/FACS), Immunofluorescence (IF)
HRP Rabbit Anti-Chicken IgY (IgG)(H+L) Antibody
Other Names:
HRP Rabbit Anti-Chicken IgY (IgG)(H+L): N/A
Host/Reactivities:
Host: Rabbit
Purification:
Affinity Purification
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC)
Anti-Mouse IgM Monoclonal Antibody (PE/Cyanine5 Conjugated) [RMM-1]
Gene Names:
IgM ; Ighm; muH; Igh6; Igh-6; Igh-M; AI326478
Other Names:
Mouse IgM; Immunoglobulin heavy constant mu; Immunoglobulin M
Host/Reactivities:
Host: Rat / Reactivity: Mouse
Isotype/Clone:
Isotype: IgG2a, kappa / Clone RMM-1
Applications:
Flow Cytometry (FC/FACS)
Anti-Mouse IgM Monoclonal Antibody (PE/Cyanine5 Conjugated) [RMM-1]
Gene Names:
IgM ; Ighm; muH; Igh6; Igh-6; Igh-M; AI326478
Other Names:
Mouse IgM; Immunoglobulin heavy constant mu; Immunoglobulin M
Host/Reactivities:
Host: Rat / Reactivity: Mouse
Isotype/Clone:
Isotype: IgG2a, kappa / Clone RMM-1
Applications:
Flow Cytometry (FC/FACS)
FITC-Donkey Anti Rabbit IgG (H+L)
Host/Reactivities:
Host: Donkey
Applications:
Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC)
HRP Rabbit Anti-Goat IgG (H+L) Antibody
Other Names:
HRP Rabbit Anti-Goat IgG (H+L): N/A
Host/Reactivities:
Host: Rabbit
Purification:
Affinity Purification
Applications:
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC)
Cy3-Rabbit Anti-Goat IgG (H+L) Antibody
Other Names:
Cy3 Rabbit Anti-Goat IgG (H+L): N/A
Host/Reactivities:
Host: Rabbit
Purification:
Affinity Purification
Isotype/Clone:
Isotype: Cy3
Applications:
Immunocytochemistry (ICC), Flow Cytometry (FC/FACS)
Anti-Human IgM Monoclonal Antibody (AF488 Conjugated) [MHM-88]
Other Names:
Human IgM; Immunoglobulin heavy constant mu; IGHM; Immunoglobulin M; Ig mu chain C region
Host/Reactivities:
Host: Mouse / Reactivity: Human
Isotype/Clone:
Isotype: IgG1, kappa / Clone MHM-88
Applications:
Flow Cytometry (FC/FACS)
FITC Goat Anti-Mouse IgG (H+L) Antibody
Other Names:
FITC Goat Anti-Mouse IgG (H+L): N/A
Host/Reactivities:
Host: Mouse
Purification:
Affinity Purification
Applications:
Immunocytochemistry (ICC), Flow Cytometry (FC/FACS)