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Secondary Antibodies

abinScience secondary antibodies are optimized for Western blot, IHC/IF, ELISA and flow cytometry. Choose cross-adsorbed and highly cross-adsorbed specificities to minimize species cross-reactivity, Fc-specific or H+L binding for assay needs, and F(ab')2 fragments for FcR-rich tissues. Conjugates include HRP/AP, Alexa Fluor 488/555/594/647, FITC, PE, APC and biotin. Each lot provides recommended dilutions, application images and CoA for consistent performance. RUO

Why Researchers Choose abinScience

Cross-Adsorbed Reduce background in multi-species assays
Specificity Options Fc-specific, H+L, subclass-specific (IgG1/IgG2a…)
F(ab')2 Fragments For IHC/IF in Fc receptor–rich samples
Bright Conjugates HRP/AP, AF488/555/594/647, FITC, PE, APC, biotin
Lot-Specific QC Images, dilution guidance, CoA

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Selection Guide

  1. Match Host & Isotype: Select secondary against your primary’s host (e.g., anti-rabbit IgG); choose H+L for broad binding or Fc-specific to avoid light-chain cross-detection.
  2. Cross-Adsorption Level: Use cross-adsorbed for multi-species panels; highly cross-adsorbed for lowest background in tissues with mixed species proteins.
  3. Fragment Choice: F(ab')2 reduces Fc-mediated binding in IHC/IF and improves penetration; whole IgG maximizes signal in WB/ELISA.
  4. Conjugate: HRP/AP for WB/ELISA; AF488/555/594/647 for IF; PE/APC for Flow; consider brightness, laser/filter compatibility and spectral overlap.
  5. Dilution & Storage: Follow lot-specific recommendations; protect fluorophores from light and avoid repeated freeze–thaw.

Applications

WB with HRP/AP chemiluminescent colorimetric detection
IHC/IF high-contrast imaging with bright, stable fluorophores
Flow cytometry panels with minimal spillover
ELISA with high S/N and broad dynamic range

Documents & QC

Datasheet with cross-adsorption matrix, isotype, and specificity
Recommended dilutions by application and representative images
Lot-specific CoA; storage and stability instructions

FAQs

H+L vs Fc-specific—how do I choose?

H+L recognizes both heavy and light chains for robust detection; Fc-specific avoids light-chain cross-reactivity (e.g., when multiple primaries share light chains) and reduces background in certain assays.

When should I use F(ab')2 fragments?

Use F(ab')2 in IHC/IF when Fc receptor–bearing cells (e.g., macrophages) cause background, or when better tissue penetration is desired.

What does “cross-adsorbed” mean?

Secondary antibodies are adsorbed against serum proteins from non-target species to minimize cross-reactivity; “highly cross-adsorbed” further reduces off-target binding in multi-species samples.

Which fluor should I pick for Flow/IF?

Match lasers/filters (e.g., AF488 for 488 nm, AF647 for 633–640 nm); consider dye brightness and panel spillover to maintain resolution.

Do I need a secondary-only control?

Yes—include a no-primary/secondary-only control to assess non-specific binding and set gates/background.

RUO disclaimer

All antibodies are for Research Use Only and not for diagnostic procedures.

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