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Alkaline Phosphatase conjugation / labeling in < 4 hrs with 30 secs hands-on time using Alkaline phosphatase Conjugation Kit - Lightning-Link® ab102850. 100% antibody recovery.


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What's included?

3 x 10 µg
Components
AP-Mix
3 x 10 µg
Modifier reagent
1 x 200 µL
Quencher reagent
1 x 200 µL

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Alkaline Phosphatase conjugation / labeling in < 4 hrs with 30 secs hands-on time using Alkaline phosphatase Conjugation Kit - Lightning-Link® ab102850. 100% antibody recovery.

Storage

Shipped at conditions

Ambient - Cannot Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

An alternative is now in stock. The alkaline phosphatase enzyme used in this kit are produced recombinantly (*in vitro*) for high batch-to-batch consistency and long term security of supply and scalability. The conjugation protocol, buffer requirements, storage conditions, and the product sizes are identical. 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 . 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 . Use the  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 sizeRecommended maximum
amount of antibody
Maximum antibody
volume1
3 x10 μg3 x 10 μg3 x 10 μL
100 μg1 x 100 μg1 x 100 μL
3 x 100 μg3 x 100 μg3 x 100 μL
1 mg1 x 1 mg1 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% Tris10.1% BSA50% glycerol
0.1% sodium azidePBSPotassium phosphate
Sodium chlorideHEPESSucrose
Sodium citrateEDTATrehalose

1 Tris buffered saline is almost always ≤ 50 mM / 0.6%

Incompatible buffer constituents

ThiomerosalProclinGlycine
ArginineGlutathioneDTT

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

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Western blot - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850), expandable thumbnail
    Image from Feng, Linyuan, et al., PLoS pathog., 14(1): e1006867; doi: 10.1371/journal.ppat.1006867. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Western blot - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850)

    Western blot - Alkaline phosphatase Conjugation Kit;- Lightning-Link.

    Feng, Linyuan, et al used Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850) as part of examining the interaction between UL23 and Nmi identified by coimmunoprecipitation. They used the kit to conjugate Alkaline phosphatase to primary antibodies for use in western blot.
    (A-B) Human U251 cells were co-transfected with a combination of two plasmids expressing FLAG- and HA-tagged proteins, and then harvested at 48 hours posttransfection. (C-D) Human U251 cells were infected with human cytomegalovirus (HCMV), a human herpesvirus, (MOI = 1) and cellular lysates were prepared at 48-72 hours postinfection. The input protein samples (80 μg) (Input) (lanes 3, 6, 9, 12, 15, 18, 21, and 24) and samples (15 μg) that were precipitated with anti-HA (IP (anti-HA)) (lanes 2, 5, 8, and 11), anti-FLAG (IP (anti-FLAG)) (lanes 1, 4, 7, and 10), anti-Nmi (lanes 13, 16, 19, and 22), anti-UL44 (lanes 14 and 17), or anti-UL23 antibodies (lanes 20 and 23), were separated on SDS-containing polyacrylamide gels, and assayed with Western blot analysis using anti-HA (anti-HA) (A), anti-FLAG (anti-FLAG) (B), anti-UL44 (anti-UL44) (C), anti-UL23 (anti-UL23) (D), and anti-Nmi (anti-Nmi) (C-D) antibodies that were directly conjugated to alkaline phosphatase using the Lightning-Link® kit ab102850, respectively.

  • Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850), expandable thumbnail
    Image from Thomas FC et al., BMC Vet Res., 11, 207. Fig 1.; doi: 10.1186/s12917-015-0533-3. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850)

    Alkaline phosphatase Conjugation Kit - Lightning-Link® labeling anti-bovine Hp antibody for ELISA.

    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).

  • Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850), expandable thumbnail
    Image from GrabiasB et al., PLoS One, 12(4): e0174229. Fig 3; doi: 10.1371/journal.pone.0174229. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850)

    Alkaline phosphatase Conjugation Kit - Lightning-Link® labeling anti P. falciparum circumsporozite protein antibody for ECL slot blot assay.

    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.

  • Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850), expandable thumbnail
    Image from Charlermroj R et al., PLoS One, 8(4): e62344. Fig 5; doi: 10.1371/journal.pone.0062344. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850)

    Alkaline phosphatase Conjugation Kit - Lightning-Link® labeling antibodies for sandwich ELISA.

    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.

  • Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850), expandable thumbnail
    Image from Thaitrong N et al., PLoS One, 8(12): e83231. Fig .; doi: 10.1371/journal.pone.0083231. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

    Conjugation - Alkaline phosphatase Conjugation Kit - Lightning-Link® (ab102850)

    Alkaline phosphatase Conjugation Kit - Lightning-Link® labeling MPC and MYSV6 polyclonal antibodies for microfluidic sandwich ELISA.

    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).

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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