Product nameAlkaline phosphatase Conjugation Kit - Lightning-Link®
Alkaline Phosphatase Conjugation Kit / Alkaline Phosphatase Labeling Kit ab102850 uses a simple and quick process for alkaline phosphatase 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 Alkaline Phosphatase using this kit:
- add modifier to antibody and incubate for 3 hrs
- add quencher and incubate for 30 mins
The alkaline phosphatase 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.
This product is manufactured by Expedeon, an Abcam company, and was previously called Lightning-Link® Alkaline Phosphatase Labeling Kit. 702-0005 is the same as the 100 µg size. 702-0010 is the same as the 3 x 100 ug size. 702-0030 is the same as the 3 x 10 ug size. 702-0015 is the same as the 1 mg size.
Amount and volume of antibody for conjugation to Alkaline Phosphatase
Kit size Recommended maximum
amount of antibody
3 x10 µg 3 x 10 µg 3 x 10 µL 100 µg 1 x 100 µg 1 x 100 µL 3 x 100 µg 3 x 100 µg 3 x 100 µL 1 mg 1 x 1 mg 1 x 1 mL
1 Ideal antibody concentration is 1mg/ml. 0.5 - 1 mg/ml can be used if the maximum antibody volume is not exceeded. Antibodies > 1 mg/ml or < 0.5 mg/ml should be diluted /concentrated.
Buffer Requirements for Conjugation
Buffer should be pH 6.5-8.5.
Compatible buffer constituents
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.
50mM / 0.6% Tris1 0.1% BSA 50% glycerol 0.1% sodium azide PBS Potassium phosphate Sodium chloride HEPES Sucrose Sodium citrate EDTA Trehalose
1 Tris buffered saline is almost always ≤ 50 mM / 0.6%
Incompatible buffer constituents
Thiomerosal Proclin Glycine Arginine Glutathione DTT
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.
Tested applicationsSuitable for: Conjugationmore details
Storage instructionsStore at -20°C. Please refer to protocols.
Components 1 mg 100 µg 3 x 10 µg 3 x 100 µg ab274118 - AP-Mix 1 x 1mg 1 x 100µg 3 x 10µg 3 x 100µg ab274119 - Modifier reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl ab274296 - Quencher reagent 1 x 200µl 1 x 200µl 1 x 200µl 1 x 200µl
Our Abpromise guarantee covers the use of ab102850 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Conjugation||Use at an assay dependent dilution.|
This illustration demonstrates a general procedure and will slightly vary dependent on the conjugate used.
Thomas FC et al used ab102850 as part of examining the toxicity of p17 protein assemblies.
They used the kit to conjugate Alkaline phosphatase to anti-bovine Hp antibody for use in ELISA.
Boxplot showing Hp concentration (μg/ml) in two SCC categories of composite milk samples * indicates an extreme value (values greater than 3 interquartile range (IQR) away from 25th or 75th percentile); IQR = 3rd quartile -1st quartile (represented by the height of the box). ° indicates an outlier value (values greater than 1.5 interquartile range (IQR) away from 25th or 75th percentile); IQR = 3rd quartile -1st quartile (represented by the height of the box).
Grabias B et al. used ab102850 as part of developing a rapid detection of Plasmodium falciparum infection in mosquitoes.
They used the kit to conjugate Alkaline phosphatase to anti-Plasmodium falciparum circumsporozite protein antibody for use in ECL slot blot assay.
Evaluation of whether conjugation of primary mAb 2A10 with alkaline phosphatase (1°-AP) enhanced sensitivity for the detection of Pfoocyst prepared from mosquito lysates when compared to the use of AP-conjugated secondary antibody (2°-AP). Detection limits were compared for each protocol by fitting the band intensities of serially diluted oocysts to a Michaelis-Menten regression curve and establishing a cutoff intensity threshold of mean + 2 SD from unfed mosquito specimens run on the same blot. Labeled primary antibody displayed overall higher band intensities across the range of oocyst dilutions examined and achieved lower limits of detection than the typical sandwich antibody format (0.009 oocyst versus 0.02 oocyst, respectively). The removal of an additional antibody incubation step also contributed to an overall shorter assay time in the newly developed slot blot protocol.
Charlermroj R et al. used ab102850 to run a sandwich ELISA and compare its sensitivity with a microsphere immunoassay based on Luminex using the same set of antibodies.
Thaitrong N et al. used ab102850 to run a microfluidic sandwich ELISA for Acidovorax citrulli (Ac), watermelon silver mottle virus (WSMoV), and melon yellow spot virus (MYSV) screening. Nine different conditions for each disease panel were tested on the microfluidic platform using combinations of three concentrations of capture Ab (11E5, 2D6, and 5E7) and three concentrations of detection Ab (MPC-AP, MYSV6-AP).
ab102850 has been referenced in 14 publications.
- Leuzy A et al. Longitudinal tau and metabolic PET imaging in relation to novel CSF tau measures in Alzheimer's disease. Eur J Nucl Med Mol Imaging 46:1152-1163 (2019). PubMed: 30610252
- Mechaly A et al. Inhibition of Francisella tularensis phagocytosis using a novel anti-LPS scFv antibody fragment. Sci Rep 9:11418 (2019). PubMed: 31388083
- Simões PBA et al. Pilot study into milk haptoglobin as an indicator of udder health in heifers after calving. Res Vet Sci 116:83-87 (2018). PubMed: 28601196
- Feng L et al. Human cytomegalovirus UL23 inhibits transcription of interferon-? stimulated genes and blocks antiviral interferon-? responses by interacting with human N-myc interactor protein. PLoS Pathog 14:e1006867 (2018). PubMed: 29377960
- Lo Passo C et al. Molecular characterization of two sub-family specific monoclonal antibodies to meningococcal Factor H binding protein. Heliyon 4:e00591 (2018). PubMed: 29644339