HRP antibody labeling protocol

HRP antibody labeling protocol

Direct detection is often a preferred method within these applications to avoid cross-species reactivity and to eliminate additional laborious wash and separation steps, especially in time-consuming protocols. However, directly conjugating HRP to the antibody or protein of choice can be difficult and labor-intensive if using traditional methodology.

Our Lightning-Link HRP antibody labeling kit allows the direct conjugation of your antibody, protein, or peptide to HRP for use in any application. The kit only requires 30 seconds of hands-on time, and conjugates are ready to use after only 3 hours. Furthermore, there is no need for extra wash or separation steps post conjugation.

​Directly label your primary antibody with HRP

​​​Horseradish peroxidase (HRP) is a 44 kDa glycoprotein with 6 lysine residues, which can be conjugated to antibodies and proteins for use in a variety of applications.

The enzyme label can be visualized through chromogenic reactions. For example, diaminobenzidine (DAB) in the presence of hydrogen peroxide (H202) is converted into a water-insoluble brown pigment. Other substrates that can be used to measure horseradish peroxidase activity include ABTS, TMB, and TMBUS.

HRP is a popular detection label used in research. Antibody-HRP conjugates are commonly used in ELISA, IHC, and western blotting. HRP can be conjugated to the primary antibody for direct detection or secondary antibody for indirect detection.

 

hrp conjugated antibody
hrp conjugated antibody

Benefits of using Lightning-Link HRP

  • Quick and easy – 30 seconds of hands-on time
  • Adjustable ratios of antibody to HRP, between 1:1 and 1:4
  • No separation steps involved, recover 100% of your antibody or protein
  • Potentially more sensitive when compared to traditional methods – so less antibody required (see figure 1 below)
  • Label from as little as 10 µg – scale up to a gram or more.

Abcam’s HRP secondaries advantages are:

  • High sensitivity – detect low-abundant targets with high signal-to-noise ratios due to an optimal number of HRP molecules per antibody.
  • Robustness – can be used at dilutions ranging from 1/2,000 to 1/20,000 and provide high specificity and minimal background.
  • Stability – they are supplied in a liquid format that is ready-to-use and can be stored at 4°C.
  • Flexibility – compatible with chromogenic, fluorogenic and chemiluminescent substrates.
  • Cost-effective – use less conjugate per assay and perform more blots.
hrp conjugated antibody
hrp conjugated antibody

Anti-mouse HRP secondary antibodies

Our secondary antibody selection guide will help you choose the most appropriate secondary antibody for your experiment. Read it now to learn more about immunoglobulins classes and subclasses, preadsorbed secondary antibodies and F(ab) or F(ab’)2 fragment secondary antibodies.

Host Non-preadsorbed* Preadsorbed*
Goat Anti-mouse HRP (IgG H&L) Yes
Anti-mouse IgG Fc HRP Yes
Anti-mouse IgG1 heavy chain HRP Yes
Anti-mouse IgG2a heavy chain HRP Yes
Anti-mouse IgG2b heavy chain HRP Yes
Anti-mouse IgG2c heavy chain HRP Yes
Anti-mouse IgG3 heavy chain HRP Yes
Anti-mouse IgM mu chain HRP Yes
Anti-mouse IgA alpha chain HRP Yes
Anti-mouse IgG+IgM HRP (H&L) Yes
Anti-mouse IgG+IgM+IgA HRP (H&L) Yes
F(ab) Anti-mouse HRP (IgG H&L) No
F(ab’)2 Anti-mouse HRP (IgG H&L) Yes
F(ab’)2 Anti-mouse IgG Fc HRP Yes
F(ab’)2 Anti-mouse IgG F(ab’)2 HRP Yes
F(ab’)2 Anti-mouse IgM mu chain HRP Yes
F(ab’)2 Anti-mouse IgG+IgM+IgA HRP (H&L) Yes
Rabbit Anti-mouse HRP (IgG H&L) Yes
F(ab’)2 Anti-mouse HRP (IgG H&L) Yes
Donkey Anti-mouse HRP (IgG H&L) Yes
F(ab’)2 Anti-mouse HRP (IgG H&L) Yes

* Preadsorbed (cross-adsorbed) antibodies go through an extra purification step to minimize species cross-reactivity and background. The process involves passaging of the antibody solution through a column containing immobilized serum proteins from potentially cross-reactive species. Nonspecific cross-reactive secondary antibodies remain bound to the proteins in the column, while highly specific secondary antibodies are recovered from the flow-through.

 

Biotin [HRP]

DAG3197-HRP 1mg
EUR 429

Vitamine B9 [HRP]

DAGDT1440-HRP 1mg
EUR 1105

Vitamine B12 [HRP]

DAGT5413-HRP 1mg
EUR 975

HRP-IgG Conjugation Kit

55R-HRP 1 mg
EUR 705
Description: HRP conjugation kit for all types of IgG

Anti-Per.C6Ð’ Affinity Purified HRP, Goat

PC118AF-HRP 1 mg
EUR 4069
Description: Anti-Per.C6Ð’ Affinity Purified HRP, Goat by Cygnus Technologies is available in Europe via Gentaur.

Goat Anti-Rat IgG1-HRP Conjugate

50121-HRP 0.5 ml
EUR 225

Goat Anti-Rat IgG2a-HRP Conjugate

50122-HRP 0.5 ml
EUR 225

Goat Anti-Rat IgG2b-HRP Conjugate

50123-HRP 0.5 ml
EUR 225

Goat Anti-Rat IgG2c-HRP Conjugate

50124-HRP 0.5 ml
EUR 225

Monoclonal Anti-Rat IgG1-HRP Conjugate

50126-HRP 0.5 ml
EUR 286

Rabbit Anti-Bird IgM-HRP Conjugate

90521-HRP 100 ul
EUR 347

Rabbit Anti-Salmonella IgG-HRP conjugate

SLM11-HRP 0.5 ml
EUR 347

Sambucus nigra Lectin (SNA)-HRP conjugate

SNA15-HRP 0.5 ml
EUR 286

Lens culinaris Lectin (LCA)-HRP conjugate

LCA15-HRP 0.5 ml
EUR 286

Monoclonal Anti-Poly His-HRP conjugate

HISP12-HRP 100 ul
EUR 347

Concanavalin A Lectin (ConA)-HRP conjugate

CONA15-HRP 0.5 ml
EUR 286

Aleuria Aurantia Lectin (Aal)-HRP conjugate

AAL15-HRP 0.5 ml
EUR 225

Goat Anti-Donkey IgG (Fc)-HRP conjugate

80950-HRP 0.5 ml
EUR 225

Goat Anti-Human Transferrin IgG-HRP conjugate

TF11-HRP 100 ug
EUR 347

Goat Anti-Mouse Transferrin IgG-HRP conjugate

TF14-HRP 100 ug
EUR 347

Wheat germ agglutinin Lectin (WGA)-HRP conjugate

WGA15-HRP 0.5 ml
EUR 286

Chicken Anti-Protein-A IgG-HRP conjugate

PRTA13-HRP 100 ul
EUR 347

Recombinant (E.Coli) purified Protein G-HRP Conjugate

PRTG15-HRP 1 ml
EUR 225

Monoclonal Anti-Phosphotyrosine (PY20) IgG-HRP conjugate

PTYR12-HRP 100 ul
EUR 408

Goat Anti-human lactoferrin IgG-HRP Conjugate

LTF11-HRP 0.5 ml
EUR 445

Goat Anti-bovine lactoferrin IgG-HRP conjugate

LTF13-HRP 0.5 ml
EUR 445

Rabbit Anti-bovine Catalase IgG-HRP Conjugate

CATB11-HRP 100 ul
EUR 469

Sheep Anti-rat albumin IgG-HRP Conjugate

ALBR12-HRP 100 ul
EUR 347

Goat Anti-Turkey IgG (H+L)-HRP conjugate

60600-HRP 0.5 ml
EUR 225

Rabbit Anti-Donkey IgG (H+L)-HRP conjugate

80920-HRP 1 ml
EUR 213

Goat Anti-Human Plasma Fibrinogen IgG-HRP Conjugate

FIBN17-HRP 100 ul
EUR 408

Goat Anti-Mouse IgG (Fc) IgG-HRP Conjugate

40151-HRP 1 ml
EUR 225

Rabbit Anti-Human alpha-Tubulin IgG-HRP Conjugate

TUBA13-HRP 100 ul
EUR 432

Ricinus communis (castor bean) Agglutinin (RCA120)-HRP conjugate

RCA15-HRP 0.5 ml
EUR 286

Goat Anti-Human plasminogen protein IgG, HRP conjugate

PLMN12-HRP 100 ul
EUR 408

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