Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

Secondary Antibodies

Search our portfolio of over 7,000 high quality purified secondary antibodies for the detection of primary antibodies in various applications, e.g. flow cytometry, cell imaging, Western blots, or CUT&Tag. We offer conjugated secondaries with a wide range of fluorescent and non fluorescent conjugates such as FITC, HA, HRP, Atto conjugates, DyLight, HiLyte and others. Discover how our PhD customer support can help you in case of any question.

Secondary Antibodies - The Right Product for Every Application

The lists below may help you to search our secondary antibodies. Follow the links to the prefiltered search, where you can apply additional filters. In case we can help you to find the right product, contact our scientific customer support via chat or phone.

Reactivities:

Fluorescent Conjugates:

Fluophore Excitation Peak (nm) Emission Peak (nm) Emission Color
DyLight 405 400 421 Image
Alexa Fluor 405 401 421
DyLight 350 352 435
Alexa Fluor 350 346 442
Alexa Fluor 488 493 519
FITC 492 520
ATTO 488 501 523
Cy3 550 570
Alexa Fluor 594 550 570
Phycoerythrin (PE) 565 578
ATTO 594 591 616
APC 650 661
Alexa Fluor 647 651 667
Cy5 650 670
PerCp 490 675
Cy5.5 675 694
Alexa Fluor 700 702 723
Cy7 778 754
DyLight 800 771 796
Alexa Fluor 790 784 814

Conjugates:

Secondary by Application:

Featured Secondary Antibodies

Product
Application
Cat. No.
Quantity
Datasheet
Application ELISA, IHC, WB, CUT&Tag, CUT&RUN
Cat. No. ABIN101961
Quantity 200 μL
Datasheet Datasheet
Application IA
Cat. No. ABIN101990
Quantity 2 mg
Datasheet Datasheet
Application ELISA, IHC, WB, DB, IEM
Cat. No. ABIN964977
Quantity 100 μg
Datasheet Datasheet

Secondary Antibody Selection Guide

Secondary antibodies are usually designed to work in specific applications. For any given application several similar secondary antibodies may be available. While primary antibodies are raised to detect a certain antigen secondary antibodies are designed to detect a primary antibody.

Selection of Secondary Antibodies

  • Determine in which species the primary antibody has been raised
  • Determine what subclass the primary antibody belongs to
  • Select a suitable marker for the secondary antibody
  • Make sure the secondary antibody works under the specific application condition
  • antibodies-online.com offers a broad range of various secondaries
  • Filter our secondary antibodies by species, conjugate, Ig class

Antibody Markers

Secondary antibodies are usually designed to work in specific applications. For any given application several similar Secondary antibodies may be available. While primary antibodies are raised to detect a certain antigen secondary antibodies are designed to detect a primary antibody.

In order to do so, they are usually labeled with markers, or conjugates in order to detect and visualize the primary antibodies, most often via colorimetric detection or emission of a light signal. Secondary antibodies can be either enzyme labeled (peroxidase, alkaline phosphatase), fluorescence labeled (e.g. FITC, Alexa-Fluor, Qdot) or biotin conjugated. Peroxidase is economical, rapid and a more stable enzyme, while the alkaline phosphatase on the other hand is considered more sensitive than peroxidase particularly when colorimetric detection is used. Fluorescent labeled antibodies are often used for double or multiple staining methods. Generally, any one of a range of markers or labels can be chosen, depending on the application and purpose of the investigation. In case of parallel detection of different antigens in the same sample (like on an IHC sample) one needs to select different monoclonal antibody classes (in order to avoid interference of secondary binding) and different markers in order to distinguish the antigen signals.

Custom Antibody Services

Custom Antibody Services

Benefit from combined expertise of antibodies-online and our Partner Rockland Immunochemicals, Inc. Collaborative, experienced, and conducted completely in our own facilities, our custom antibody production services can drive your project from the design and synthesis of your antigen through to production and characterization of your antibody.

Benefit from combined expertise of antibodies-online and our Partner Rockland Immunochemicals, Inc. Collaborative, experienced, and conducted completely in our own facilities, our custom antibody production services can drive your project from the design and synthesis of your antigen through to production and characterization of your antibody.

Learn more

Antibody Classes

When choosing a secondary antibody, first of all the suitable secondary antibody for recognition of the correct primary antibody like IgG, IgM, IgE, IgD or IgA needs to be selected. The significant difference between these isoforms are their heavy chains. They are denominated with Greek letters which are the basis for the antibodies' names: gamma, mu, epsilon, delta and alpha. Each antibody has two light chains (in mammals kappa or lambda) which also must be taken in to account for some applications (obviously if the secondary antibody is to bind not the Fc part or other regions of the heavy chains).

For the selection of the correct secondary antibody the specific subtype must be observed in most cases, i.e. if the subclass is IgG1, IgG2a, IgG2b, IgG2c, IgG3 and so on. However, general anti-IgG secondary antibodies may recognize various subtypes. Additionally, secondary antibodies can bind all parts of an IgG molecule (heavy and light chains, H+L), or only the Fab or Fc region, or only the gamma chain. Still others are specific for IgM heavy chains, or the lambda or kappa light chains common to all immunoglobulins (IgG, IgA, IgD, IgE, and IgM).

The secondary antibody will usually belong to the classes IgG or IgM.

Antibody Fragments

A secondary antibody can further be defined as an antibody which binds to primary antibodies or antibody fragments. Secondary antibodies may be polyclonal or monoclonal, even though they most often derive from polyclonal sources and are affinity purified against the epitope they have been developed for. Secondary antibodies may hence be specific for whole Ig molecules or fragments of primary antibodies, such as the Fc or Fab regions.

Fc and Fab fragments can be generated with the enzyme papain that can be used to cleave an immunoglobulin monomer into two Fab fragments and an Fc fragment. Proteolytic enzyme pepsin cleaves below the hinge region, forming a F(ab')2- and a pFc' fragment. The F(ab')2 fragment can be split into two Fab' fragments by reduction reaction.

For certain applications, antibody fragments are used as the primary antibody in order to obtain the best results. In that case a secondary antibody is used that is specific to the primary antibody fragment. For example, in immunohistochemistry (IHC), background noise may be reduced if a Fab fragment of the primary antibody is used instead of the whole antibody molecule, because binding to endogenous Fc molecules in the sample does not occur with Fab-specific secondary antibodies.

Another situation occurs, when the secondary antibody itself is a fragment. For example, if working with tissues or cells that have Fc receptors (thymus, spleen, blood, hematopoietic cells, leukocytes, B cells, etc.) a F(ab')2 fragment secondary antibodies can be selected in order to avoid Fc receptor binding of the secondary antibody on the cells.

Immunoglobulin Isoforms

Good Specificity and High Sensitivity

The criteria for selecting the right secondary antibody are extremely good specificity and high sensitivity. antibodies-online offers you more than 5000 secondary antibodies validated for the most varied applications, such as ELISA, flow cytometry, western blot, ICC, IHC and many more. Our product portfolio covers all common host organisms and reactivity types. But you’ll also find secondary antibodies from more rarely used hosts, such as apes, dogs, Syrian hamsters and others.

Choose from a multitude of conjugated secondary antibodies, for example products combined with TRITC, FITC, Cy, Dylight and Alexa dyes. Still can’t find the right product? Then contact our customer support team via chat, phone or e-mail.

Featured Secondary Antibodies

Product
Reactivity
Source
Application
Cat. No.
Reactivity Rabbit
Source Guinea Pig
Application Immunoassay (IA)
Cat. No. ABIN101961
Reactivity Rabbit
Source Goat
Application Immunomicroscopy (IM), FLISA, Western Blotting (WB)
Cat. No. ABIN101988
Reactivity Rabbit
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN101990
Reactivity Goat
Source Rabbit

Cat. No. ABIN1720731
Reactivity Rabbit
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN964980
Reactivity Rabbit
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN964982
Reactivity Rabbit
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN964987
Reactivity Rabbit
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN964991
Reactivity Human
Source Mouse
Application Flow Cytometry (FACS)
Cat. No. ABIN94404
Reactivity Mouse
Source Goat
Application
Cat. No. ABIN6699005
Reactivity Rabbit
Source Goat
Application Immunomicroscopy (IM), FLISA, Western Blotting (WB)
Cat. No. ABIN964975
Reactivity Goat
Source Rabbit
Application Immunoassay (IA)
Cat. No. ABIN1043943
Reactivity Mouse
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN964962
Reactivity Rabbit
Source Goat
Application Immunomicroscopy (IM), FLISA, Western Blotting (WB)
Cat. No. ABIN101991
Reactivity Rabbit
Source Goat
Application Western Blotting (WB), Dot Blot (DB), ELISA, Immunohistochemistry (IHC)
Cat. No. ABIN101985
Reactivity Rabbit
Source Goat
Application Immunomicroscopy (IM), Flow Cytometry (FACS)
Cat. No. ABIN129765
Reactivity Human
Source Mouse
Application ELISA
Cat. No. ABIN135676
Reactivity Rabbit
Source Goat

Cat. No. ABIN2176507
Reactivity Mouse
Source Goat
Application Immunoassay (IA)
Cat. No. ABIN1043983
Reactivity Mouse
Source Goat
Application Flow Cytometry (FACS), Immunohistochemistry (IHC)
Cat. No. ABIN376045
Reactivity Goat
Source Chicken
Application
Cat. No. ABIN7273065
Reactivity Chicken
Source Chicken
Application
Cat. No. ABIN7273054
Reactivity Chicken
Source Chicken
Application
Cat. No. ABIN7273051
Reactivity Chicken
Source Mouse
Application Flow Cytometry (FACS), Immunohistochemistry (IHC)
Cat. No. ABIN376731
Reactivity Rabbit
Source Goat
Application
Cat. No. ABIN6699118
Reactivity Rabbit
Source Donkey
Application
Cat. No. ABIN6699117
Reactivity Rabbit
Source Donkey
Application
Cat. No. ABIN6699106
Reactivity Rabbit
Source Goat
Application
Cat. No. ABIN6699089
Reactivity Mouse
Source Goat

Cat. No. ABIN2176530
Reactivity Rat
Source Goat

Cat. No. ABIN2176434
Reactivity Rabbit
Source Donkey

Cat. No. ABIN2176495
Reactivity Rabbit
Source Mouse

Cat. No. ABIN2176516
Reactivity Rabbit
Source Goat
Application Flow Cytometry (FACS), Immunofluorescence (IF)
Cat. No. ABIN612374
Reactivity Rabbit
Source Goat
Application ELISA, Western Blotting (WB), Immunohistochemistry (IHC)
Cat. No. ABIN375875
Reactivity Human
Source Goat
Application Flow Cytometry (FACS), Immunofluorescence (IF)
Cat. No. ABIN612018
You are here:
Support