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.

 

HRP antibody

20C-CR7015GP 500 ul
EUR 52
Description: Goat polyclonal HRP antibody

HRP antibody

20R-3010 100 µg
EUR 697
Description: Rabbit polyclonal HRP antibody

HRP Antibody

E10-20162 100μg/100μl
EUR 225
Description: Available in various conjugation types.

HRP Antibody

20-abx134340
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  • 100 ul
  • 200 ul
  • 30 ul

HRP Antibody

abx015753-100ul 100 ul
EUR 493.2

HRP Antibody

abx015753-1mg 1 mg
EUR 362.5

GST HRP Antibody

48050 100ul
EUR 429

GST HRP Antibody

48050-100ul 100ul
EUR 399.6

GST HRP Antibody

48050-50ul 50ul
EUR 286.8

Prion -HRP, Antibody

GWB-32B00A 0.25 ml Ask for price

Ferritin -HRP, Antibody

GWB-A83A47 0.5 mg Ask for price

Peroxidase (HRP) Antibody

GWB-93765A 50 mg Ask for price

anti- HRP 2 antibody

FNab04012 100µg
EUR 606.3
Description: Antibody raised against HRP 2

I79_001071 Antibody (HRP)

20-abx333649
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

HA antibody (HRP)

60R-1086 1 ml
EUR 609.6
Description: Affinity purified Chicken polyclonal HA antibody (HRP)

P Antibody (HRP)

20-abx300952
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  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Ig Antibody (HRP)

GWB-8135B0 1 ml Ask for price

HA Antibody (HRP)

GWB-CE01D9 0.1 mg Ask for price

P Antibody (HRP)

abx300952-100g 100 µg
EUR 362.5

P Antibody (HRP)

abx300952-20g 20 µg
EUR 162.5

P Antibody (HRP)

abx300952-50g 50 µg
EUR 250

HA Tag HRP Antibody

48052 100ul
EUR 429

HA Tag HRP Antibody

48052-100ul 100ul
EUR 399.6

HA Tag HRP Antibody

48052-50ul 50ul
EUR 286.8

VWF antibody (HRP)

60R-1021 200 ug
EUR 361
Description: Sheep polyclonal VWF antibody (HRP) conjugated

tPA antibody (HRP)

60R-1069 200 ug
EUR 426
Description: Sheep polyclonal tPA antibody (HRP) conjugated

PF4 antibody (HRP)

60R-1077 200 ug
EUR 411
Description: Sheep polyclonal PF4 antibody (HRP) conjugated

PCI antibody (HRP)

60R-1080 200 ug
EUR 532
Description: Goat polyclonal PCI antibody (HRP) conjugated

RNR antibody (HRP)

60R-1112 100 ug
EUR 302
Description: Rabbit polyclonal RNR antibody (HRP)

Rho antibody (HRP)

60R-1118 100 ug
EUR 302
Description: Rabbit polyclonal Rho antibody (HRP)

PDI antibody (HRP)

60R-1269 100 ug
EUR 302
Description: Rabbit polyclonal PDI antibody (HRP)

U1A antibody (HRP)

60R-1329 100 ug
EUR 302
Description: Rabbit polyclonal U1A antibody (HRP)

TRP antibody (HRP)

60R-1344 100 ug
EUR 302
Description: Rabbit polyclonal TRP antibody (HRP)

PGD antibody (HRP)

60R-1367 100 ug
EUR 302
Description: Rabbit polyclonal PGD antibody (HRP)

SmG antibody (HRP)

60R-1418 100 ug
EUR 302
Description: Rabbit polyclonal SmG antibody (HRP)

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