Buccutite™ PolyHRP Antibody Conjugation Kit

Buccutite™ PolyHRP Antibody Conjugation Kit is designed for preparing polymeric horseradish peroxidase-labeled antibodies using complementary Buccutite™ MTA and FOL linkers.

Preactivated PolyHRP: Includes Buccutite™ FOL-activated PolyHRP and Buccutite™ MTA for antibody modification and subsequent conjugation
Two-step conjugation: Uses antibody modification followed by coupling to activated PolyHRP, with 30-minute and 60-minute incubations at room temperature
Multiple detection formats: The resulting conjugates are compatible with chromogenic, fluorogenic, and chemiluminescent HRP substrates
Applications: Supports preparation of detection antibodies for ELISA, Western blotting, immunohistochemistry, and tyramide-based signal amplification

Direct ELISA curves were generated using the polyHRP- goat-anti-mouse IgG conjugate prepared with Buccutite™ Poly-HRP Antibody Conjugation Kit (Cat No. 5518, Cat No. 5519). A 3-fold  serial diluted mouse IgG was coated onto a 96-well plate, and 100 µL GAM IgG-polyHRP conjugate (100 ng/ml) was tested using the standard ELISA method. TMB substrate solution (Cat No. 11003) was used to detect the immobilized mouse IgG with 5 min incubation and read at 450 nm.<br><br />Blue: Buccutite™ Kit 5518 or 5519<br />Red: GXM IgG PolyHRP (Vendor A)<br />Green: GXM IgG-HRP (Vendor B)

Protocol

Catalog	Size	Price	Quantity
5518	1x50 ug Labeling	Price
5519	2x50 ug Labelings	Price

Overview

Buccutite™ PolyHRP Antibody Conjugation Kit is designed for preparing antibodies conjugated to polymeric horseradish peroxidase (PolyHRP). The kit includes PolyHRP preactivated with Buccutite™ FOL, Buccutite™ MTA for antibody modification, and Reaction Buffer. Following MTA modification, the antibody is mixed with FOL-activated PolyHRP to form the conjugate through a catalyst-free reaction. The kit is available in formats for one 50 µg antibody labeling reaction, two 50 µg antibody labeling reactions, or labeling 1 mg of antibody.

The workflow uses a 30-minute antibody modification step followed by a 60-minute conjugation step at room temperature. PolyHRP provides multiple enzyme units for signal amplification, and the resulting antibody conjugates are compatible with chromogenic, fluorogenic, and chemiluminescent HRP substrates. These conjugates support ELISA, Western blotting, immunohistochemistry, and signal amplification using tyramide or Styramide™ fluorescent HRP substrates.

References

View all 11 references: Citation Explorer

Detecting cancer metastasis and accompanying protein biomarkers at single cell levels using a 3D-printed microfluidic immunoarray.

Authors:

Sharafeldin, Mohamed and Chen, Tianqi and Ozkaya, Gulsum Ucak and Choudhary, Dharamainder and Molinolo, Alfredo A and Gutkind, J Silvio and Rusling, James F

Journal:

Biosensors & bioelectronics (2021): 112681

Prostate Cancer Diagnosis in the Clinic Using an 8-Protein Biomarker Panel.

Authors:

Jones, Abby L and Dhanapala, Lasangi and Baldo, Thaísa A and Sharafeldin, Mohamed and Krause, Colleen E and Shen, Min and Moghaddam, Shirin and Faria, Ronaldo C and Dey, Dipak K and Watson, R William and Andrawis, Ramez and Lee, Norman H and Rusling, James F

Journal:

Analytical chemistry (2020)

Comparison of different methodologies and cryostat versus paraffin sections for chromogenic immunohistochemistry.

Authors:

Hira, Vashendriya V V and de Jong, Annique Loncq and Ferro, Klea and Khurshed, Mohammed and Molenaar, Remco J and Van Noorden, Cornelis J F

Journal:

Acta histochemica (2019): 125-134

The complementary role of insulin-like growth factor II mRNA-binding protein 3 (IMP3) in diagnosis of Hodgkin's lymphoma.

Authors:

Masoud, Rokia and Ibrahiem, Afaf and Tantawy, Dina and Eldosoky, Ibrahim

Journal:

Annals of diagnostic pathology (2019): 64-68

Using magnetic beads and signal amplifiers to produce short and simple immunoassays: Application to MMP-9 detection in plasma samples.

Authors:

Ben Ismail, Manel and de la Serna, Erica and Ruiz-Vega, Gisela and García-Berrocoso, Teresa and Montaner, Joan and Zourob, Mohammed and Othmane, Ali and Baldrich, Eva

Journal:

Analytica chimica acta (2018): 144-154

Usage and storage

Intended use

Research Use Only (RUO)

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Page updated on October 22, 2026

Component A: Buccutite™ FOL-Activated Poly-HRP	1 vial (200 µL/vial)
Component B: Buccutite™ MTA	1 vial (Lyophilized)
Component C: Reaction Buffer	1 Vial (20 µL)

Component A: Buccutite™ FOL-Activated Poly-HRP	2 vials (200 µL/vial)
Component B: Buccutite™ MTA	2 vials (Lyophilized)
Component C: Reaction Buffer	1 vial (40 µL)