Jackson ImmunoResearch: i migliori anticorpi secondari
Ampia gamma di anticorpi secondari ad alta AFFIDABILITA’, RIPRODUCIBILITA’ e STABILITA’.
Con oltre 7.000 anticorpi secondari Whole IgG, Fab e F(ab’)2 nel nostro database troverai sempre la giusta combinazione di coniugati specifici per il tuo saggio immunologico. La selezione comprende anticorpi secondari coniugati con horseradish peroxidase o alkaline phosphatase per Western Blotting (WB), ELISA e Immunohistochemistry (IHC) nonché anticorpi secondari coniugati per metodi basati su fluorescenza come Immunofluorescent Staining e Flow Cytometry (FACS). Le coniugazioni fluorescenti includono una selezione di fluorofori Alexa ™, Cynanin ™ (Cy2, Cy3 e Cy5) e DyLight® 405.
Come scegliere il secondario giusto?
Choosing the right affinity-purified secondary antibody for your application
Affinity-purified antibodies are isolated from antisera by immunoaffinity chromatography using antigens coupled to agarose beads. A proprietary elution process is used to dissociate antibodies from the antigen. Unconjugated affinity-purified antibodies are supplied sterile filtered in phosphate buffer without stabilizers or preservatives. Conjugated affinity-purified antibodies are freeze-dried in phosphate buffer with stabilizers and sodium azide, with the exception of horseradish peroxidase conjugates, which do not contain a preservative.
Step 1. Select from Whole IgG, F(ab’)2 Fragments, or Fab Fragments of secondary antibodies.
Affinity-purified secondary antibodies are offered in three different forms: whole IgG, F(ab’) 2 fragments, and Fab fragments.
Step 2. Select the host species of the primary antibody.
The antibodies are listed alphabetically according to the host species of the antigen (i.e. the primary antibody). For example, if the primary antibody is made in mouse, select “Anti-Mouse”. This is the target species in the product filter.
Note: Both anti-Syrian and anti-Armenian hamster secondary antibodies are listed under “Anti-Hamster”. It is important to know in which strain of hamster the primary antibody was produced in order to select the proper secondary antibody, since cross-reaction between the strains is not complete.
Step 3. Select the host species of the secondary antibody.
Selection of the host species for a secondary antibody involves many considerations, including but not limited to:
- Antibodies from some host species may not be adsorbed against cross-reacting species of interest. Chose a host species with the required adsorptions.
- Host species compatibility. Some host species may not be compatible with other species in multiple-labeling experiments. In general, all secondary antibodies should come from the same host species for multiple labeling.
- Binding to Protein A and Protein G. Rabbit antibodies bind well to Protein A and G, but goat and donkey antibodies bind better to Protein G.
- Personal preference or experience. In our experience there appears to be no species specific difference in the quality of secondary antibodies.
Step 4. Select the secondary antibody specificity under “Antibody Description”.
Note: Immunoglobulins from different species share similar structures, with similarities being related to closeness in phylogeny. Antibodies against immunoglobulins from one species may cross-react with a number of other species unless they have been specifically adsorbed against the cross-reacting species. Antibodies that have been adsorbed against other species will contain “(min X…Sr Prot)” in the antibody description.
Step 5. Select the desired probe.
Selection of fluorophores depends on:
- Instrument set-up. Examples include availability of light sources, filter sets, and detection systems.
- Degree of color separation desired for multiple labeling. For example, to achieve good color separation from Alexa Fluor® 488, choose a shorter wavelength-emitting fluorophore such as DyLight 405 and/or longer wavelength-emitting fluorphores such as Rhodamine Red-X, Alexa Fluor® 594, or Alexa Fluor® 647.
- Sensitivity required. For example, Alexa Fluor® 488 is brighter than FITC.
Effective 4-color imaging with good color separation, good photostability, and high sensitivity is possible with combinations such as DyLight 405, Alexa Fluor® 488, Rhodamine Red-X, and Alexa Fluor® 647.
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