HRP Conjugation Kit – Lightning-Link®

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Overview

Product name

HRP Conjugation Kit – Lightning-Link®

Product overview

HRP Conjugation Kit / HRP Labeling Kit ab102890 uses a simple and quick process for HRP labeling / conjugation of antibodies. It can also be used to conjugate other proteins or peptides. Learn about our antibody labeling kits and their advantages.


To conjugate an antibody to HRP using this kit:
– add modifier to antibody and incubate for 3 hrs
– add quencher and incubate for 30 mins
The HRP conjugated antibody can be used immediately in WB, ELISA, IHC etc. No further purification is required and 100% of the antibody is recovered for use.


Learn about buffer compatibility below; for incompatible buffers and low antibody concentrations, use our rapid antibody purification and concentration kits. Use the FAQ to learn more about the technology, or about conjugating other proteins and peptides to HRP.


Custom size conjugation kits up to 100 mg are available on demand. Please contact us to discuss your requirements.

Notes

This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® HRP Labeling Kit. 701-0004 is the same as the 5 mg size. 701-0003 is the same as the 5 x 1 mg size. 701-0000 is the same as the 3 x 100 ug size. 701-0030 is the same as the 3 x 10 ug size. 701-0010 is the same as the 100 µg size. 701-0002 is the same as the 1 mg size.

Amount and volume of antibody for conjugation to HRP

 Kit sizeRecommended
amount of antibody
1 
Maximum
amount of antibody
Maximum antibody
volume2
3 x 10 µg3 x 10 µg3 x 40 µg3 x 10 µL
100 µg1 x 100 µg 1 x 400 µg1 x 100 µL
3 x 100 µg3 x 100 µg 3 x 400 µg3 x 100 µL
1 mg1 x 1 mg1 x 4 mg1 x 1 mL
5 x 1 mg5 x 1 mg5 x 4 mg5 x 1 mL
5 mg1 x 5 mg1 x 20 mg1 x 5 mL

Recommended amount of antibody will give an HRP:antibody molar ratio of 4:1. Antibodies often also perform well at the maximum amount of antibody which is 1:1 molar ratio.

2 Ideal antibody concentration is 1mg/ml. 0.5 – 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 4 mg/ml or < 0.5 mg/ml should be diluted /concentrated.

 

Buffer Requirements for Conjugation

Buffer should be pH 6.5-8.5.

Buffer constituent compatibility
If a concentration is shown, then the constituent should be no more than the concentration shown. If several constituents are close to the limit of acceptable concentration, then this can inhibit conjugation.

Compatible buffer constituents
50mM / 0.6% Tris10.1% BSA50% glycerol
0.1% sodium azide2PBSPotassium phosphate
Sodium chlorideHEPESSucrose
Sodium citrateEDTATrehalose

1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
2 Sodium azide irreversibly inhibits HRP; antibodies with azide should be purified before using the HRP conjugation kit.

 

Incompatible buffer constituents
ThiomerosalProclinGlycine
ArginineGlutathioneDTT

If a constituent of the buffer containing your antibody or protein is not listed above, please check the FAQ or contact us.

Only purified antibodies are suitable for use, ie. where other proteins, peptides, or amino acids are not present: antibodies in ascites fluid, serum or hybridoma culture media are incompatible.

 

Storing and handling conjugation kits

Lyophilized Lightning-Link® components are hygroscopic.

Kits are intentionally shipped at ambient temperature with silica gel to avoid exposure to moisture. Upon receipt, store the kit frozen and protect from moisture. Before opening the outer container, allow the lyophilized components to reach room temperature to minimize condensation.

Properties

Storage instructions

Store at -20°C. Please refer to protocols.
Components100 µg1 mg3 x 10 µg5 x 1 mg3 x 100 µg1 x 5 mg
ab274112 – HRP mix1 x 100µg1 x 1mg3 x 10µg5 x 1mg3 x 100µg1 x 5mg
ab274106 – Modifier reagent1 x 200µl1 x 200µl1 x 200µl1 x 1200µl1 x 200µl1 x 1200µl
ab274296 – Quencher reagent1 x 200µl1 x 200µl1 x 200µl1 x 1200µl1 x 200µl1 x 1200µl

Research areas

  • Tags & Cell Markers
  •  
  • Epitope Tags
  •  
  • Conjugates
  • Kits/ Lysates/ Other
  •  
  • Kits
  •  
  • Conjugation Kits
  •  
  • HRP

Alternative names

  • Horseradish peroxidase

Protocols