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.
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/
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).
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 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.
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 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.
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 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).
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/
