Thermo Fisher Secondary Antibodies—F(ab')2, Cross-Adsorbed, Species-Specific

Shop Thermo Fisher (Invitrogen/Pierce) secondary antibodies for Western blot, immunohistochemistry, immunofluorescence, ELISA, and flow. Iright supplies genuine, application-validated SKUs with fast quoting and clear selectors by species, conjugate (Alexa Fluor, HRP/AP, biotin), fragment, and cross-adsorption level.

What This Collection Covers: Thermo Fisher Secondary Antibodies at Iright

Secondary antibodies bind the Fc (or H+L) region of your primary antibody and carry a detection tag to reveal the signal. On this page you’ll quickly narrow by application, species, and conjugate, then jump straight to popular SKUs—sourced from Thermo Fisher’s Invitrogen and Pierce lines—ready for your workflow.

Quick Selector: Species, Conjugates (Alexa Fluor, HRP/AP), Fragments & Cross-Adsorption

Before you compare SKUs, decide on the downstream readout and your upstream primary (host species/isotype). Next, pick the conjugate and fragment that best balances sensitivity and background for your sample type, especially in multiplex or Fc-receptor-rich tissues.

• Target species: anti-mouse, anti-rabbit, anti-human, anti-rat, etc.

• Conjugates: Alexa Fluor 488/555/594/647 for IF/flow; HRP/AP for WB/IHC/ELISA; biotin for amplification.

• Fragments: Whole IgG for routine; F(ab’)₂/Fab to reduce Fc-mediated background; Fc-specific to avoid chain cross-reactivity.

• Cross-adsorbed: Prefer highly cross-adsorbed for mixed-species/multiplex to suppress off-species binding.

Choose by Application: WB, IHC, IF, ELISA & Flow Cytometry

Match the conjugate and fragment to your assay to maximize S/N and minimize artifacts. For hard samples, start with fragment secondaries and cross-adsorbed formats, then refine dilution.

• Western blot (WB): Enzyme conjugates dominate—HRP with ECL substrates for high sensitivity; AP for durable chromogenic development. Cross-adsorbed secondaries improve band specificity in complex lysates.

• Immunohistochemistry (IHC): HRP/DAB or AP/BCIP detection; F(ab’)₂ helps reduce Fc-receptor background in spleen, liver, or macrophage-rich tissues.

• Immunofluorescence (IF): Choose bright, photostable Alexa Fluor secondaries (e.g., 488/555/647) for clean multiplex; pair dyes to instrument lasers/filters and avoid overlapping emissions.

• ELISA: HRP/AP secondaries tuned to your substrate and linear range; prioritize lot-to-lot consistency.

• Flow cytometry: Use spectrally separated Alexa Fluor dyes; block Fc receptors and consider F(ab’)₂ in sensitive cell types.

Top Categories: Alexa Fluor, HRP/AP, Cross-Adsorbed, F(ab’)₂/Fab Secondaries

Each card below links to pre-filtered results so you can land on relevant SKUs within two clicks.

• Alexa Fluor Secondaries (AF488/555/594/647) — photostable dyes for IF & flow, ideal for multiplex panels.

• HRP/AP-Conjugated Secondaries — for WB/IHC/ELISA; pair with ECL or chromogenic substrates.

• Cross-Adsorbed Formats — reduce off-species binding in multi-species samples and dual-staining.

• F(ab’)₂ / Fab Fragments — minimize Fc-mediated background and improve tissue specificity.

Comparison Table: Conjugate vs Application (Alexa Fluor/HRP/AP × WB/IHC/IF/ELISA)

Below is a quick decision grid. Use it to align conjugate chemistry to your readout; then drill into the SKUs.

Multiplex Spectra Quick Guide: AF488/555/594/647 Excitation–Emission

Use the spectra to pick non-overlapping pairs and match your lasers/filters. In classic three-color IF, AF488 + AF555 + AF647 offers clean separation on most microscopes and cytometers.
AF488: Ex ~495 nm / Em ~519 nm
AF555: Ex ~555 nm / Em ~565 nm
AF594: Ex ~590 nm / Em ~617 nm
AF647: Ex ~650 nm / Em ~668 nm.

Popular SKUs: Goat anti-Rabbit/Mouse, Alexa Fluor & HRP Secondaries

We curated high-demand Invitrogen/Pierce SKUs across dyes and enzymes. Use them as starting points, then expand via the filters above.

Product Comparison Table: Conjugate × Application (Field-Ready)

Use this compact matrix to scan attributes across conjugates and decide quickly. Populate with any additional SKUs you carry.

Multiplex Spectra Quick Sheet (Instrument-Friendly)

For panel design, map dyes to your lasers/filters and avoid emission crowding. The values below reflect the Molecular Probes handbook.

Usage Tips & FAQs: Cross-Adsorbed, F(ab’)₂, Isotype, Spectral Pairing

Choosing the right secondary often resolves background faster than changing exposure or gain. Start with species match and conjugate, then adjust fragment and cross-adsorption as needed.
FAQs

1. When should I use F(ab’)₂ or Fab fragments?
   Use fragments in Fc-receptor-rich tissues (e.g., spleen, liver) or when primary-secondary crosslinks create background. Fragments lack the Fc portion, reducing non-specific Fc binding.

2. What does “(H+L)” mean, and when do I prefer Fc-specific?
   (H+L) recognizes both heavy and light chains—maximizing capture of diverse IgGs. In multiplex with isotype-specific primaries, Fc-specific can reduce crosstalk between chains.

3. Why cross-adsorbed?
   Cross-adsorption removes off-species reactivity, crucial when using rabbit and mouse primaries together or working with human tissues. Highly cross-adsorbed donkey secondaries are the safest choice for dual-species IF.

4. WB vs IF: HRP/AP or Alexa Fluor?
   For WB, HRP with ECL is the sensitivity workhorse; AP is stable for long exposures. For IF/flow, Alexa Fluor dyes offer bright, photostable fluorescence tailored to common lasers.
5. How do I build a clean three-color IF panel?
   Start with AF488 + AF555 + AF647, confirm filters, and check for primary host conflicts. If nuclei are DAPI-stained, avoid blue-channel secondaries. Use cross-adsorbed formats in mixed species.
6. Storage & handling
   Follow the datasheet: protect from light; avoid freeze–thaw cycles; aliquot on first thaw. Check each lot’s COA for concentration/degree of labeling.

Talk to Iright

Need panel design or a one-to-one replacement for your current secondary? Tell us your primary host, applications, and instrument lasers, and we’ll shortlist compatible Thermo Fisher secondaries, bundles (substrates/antifade), and lead times. Iright can also source validated alternatives and provide COAs, bulk sizes, and cross-adsorbed options for multiplex imaging.