APC Conjugation Kit - Lightning-Link®

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - APC Conjugation Kit - Lightning-Link® (AB201807)

Haque, Nasreen S., Akaash Tuteja, and Niloufar Haque used APC Conjugation Kit - Lightning-Link^® (ab201807) as part of examining as part of examining embryoid body formation. They used the kit to conjugate APC to anti-CCR8 antibody for use in immunofluorescence.
Anti CCL1 inhibits the expression of CCR8 and FoxP3 in mouse mesenchymal stem cells (mMSCs). Untreated (5a,-c) and anti-CCL1 treated (5d,-,f) cells were subjected to immunoflourescence with antibodies against FoxP3 (green;5b and e) or CCR8 (red;5c and f).

Image from Haque, Nasreen S., Akaash Tuteja, and Niloufar Haque., PloS one 14.7 (2019): e0218944. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Schematic Diagram

Supplier Data

Schematic Diagram - APC Conjugation Kit - Lightning-Link® (AB201807)

This illustration demonstrates a general procedure of how Lightning-Link® labeling technology enables the direct labeling of antibodies or proteins.

Simply pipette your antibody or biomolecule of choice into the vial of a lyophilized mixture containing the label of interest and incubate for around 3 hours. Please see the ab201807 protocol booklet for more details.

Learn more about our Lightning-Link® conjugation kits here

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - APC Conjugation Kit - Lightning-Link® (AB201807)

Bouchery, Tiffany, et al used APC Conjugation Kit - Lightning-Link^® (ab201807) as part of examining methods for arresting hookworm development. They used the kit to conjugate APC to anti-Na-APR-1 antibody for use with live hookworms in fluorescence microscopy.
Infective L3 stage hookworms were fed in vitro for 24 hr with RBC and treated with 10 μg of 11F3-APC monoclonal antibody against Na-APR-1 or with an isotype matched-RELM-APC control antibody. Larvae were allowed to empty their digestive contents for 2 hours in fresh DMEM before internal labelling was evaluated by confocal microscopy. Data representative of 50 larvae cultured in 3 independent experiments.

Image from Bouchery, Tiffany, et al., PLoS Pathog., 14(3): e1006931; doi: 10.1371/journal.ppat.1006931. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Fluorescence Microscopy

PubMed

Fluorescence Microscopy - APC Conjugation Kit - Lightning-Link® (AB201807)

Kim, Nayoung, et al used APC Conjugation Kit - Lightning-Link^® (ab201807) as part of examining apoptosis in HIV-1-infected CD4+ primary T cells. They used the kit to conjugate APC to anti-pFOXO3a antibody for use in confocal microscopy.
Jurkat T cells expressing tTA alone, TatSF2+tTA, or TatSF2G48-R57A +tTA were stained with antibodies against PPP2R1B (first and forth columns of panels, green), pFOXO3a (second column, red), and FOXO3a (forth column, red).

Image from Kim, Nayoung, et al., PLoS Pathog., 6(9): e1001103. doi: 10.1371/journal.ppat.1001103. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - APC Conjugation Kit - Lightning-Link® (AB201807)

Flow Cytometry - APC Conjugation Kit;- Lightning-Link.

Robinson, Andrew P., et al used APC Conjugation Kit - Lightning-Link^® (ab201807) as part of characterizing oligodendroglial populations. They used the kit to conjugate APC to Mouse anti-O4 antibody, clone 81, for use in flow cytometry.
SJL/J mice were immunized with PLP139–151 and scored daily for clinical disease. A cohort of SJL/J mice was sacrificed, and spinal cords were analyzed by flow cytometry (n = 5). (A) Cells were distinguished from debris by forward and side scatter then singlet cells were gated. Live cells were gated by dead cell exclusion, and CNS resident cells were identified as CD45− or CD45low. (B) Oligodendroglial cells were defined by double positive staining : A2B5+PDGFRα+ early OPCs, A2B5+NG2+ intermediate OPCs, NG2+O4+ late OPCs, O4+MOG+ pre-myelinating oligodendrocytes, and GALC+MOG+ mature oligodendrocytes.

Image from Robinson, Andrew P., et al., PloS one, 9(9):e107649. doi: 10.1371/journal.pone.0107649. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - APC Conjugation Kit - Lightning-Link® (AB201807)

Flow Cytometry - APC Conjugation Kit- Lightning-Link.

Drachsler, Moritz, et al used APC Conjugation Kit - Lightning-Link^® (ab201807) as part of examining expression of CD95. They used the kit to conjugate APC to anti-human CD95 for use in flow cytometry.
The graph shows the CD95 expression in seven patient tumor samples.

Image from Drachsler, Moritz, et al., Cell death & disease 7.4 (2016): e2209-e2209. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Flow Cyt

PubMed

Flow Cytometry - APC Conjugation Kit - Lightning-Link® (AB201807)

Dunne, Margaret R., et al used APC Conjugation Kit - Lightning-Link^® (ab201807) as part of examining coeliac disease. They used the kit to conjugate APC to Vd3 antibody for use in flow cytometry.
Dotplots show flow cytometry data for representative control, treated and untreated coeliac donors.

Image from Dunne, Margaret R., et al., PLoS One, 8(10): e76008, doi: 10.1371/journal.pone.0076008. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - APC Conjugation Kit - Lightning-Link® (AB201807)

Image from Kalis, M et al., PLoS One, 6(12):e29166. Fig 5.; doi: 10.1371/journal.pone.0029166. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - APC Conjugation Kit - Lightning-Link® (AB201807)

APC Conjugation Kit - Lightning-Link® labeling OVA and OVA-CRT for in-gel Fluorescence and Flow cytometry.

Del Cid N et al. used ab201807 as part of examining antigen cross-presentation.

They used the kit to conjugate APC to ovalbumin (OVA) and ovalbumin-calreticulin fusion protein (OVA-CRT) for use in in-gel fluorescence and flow cytometry.
OVA-CRT and OVA were labeled with allophycocyanin. (C) Labeling intensity was determined by fluorescence imaging of the proteins after separation by SDS-PAGE (inset). Fluorescence intensity was quantified for the indicated proteins. (D) Binding of fluorescent proteins to BMDC was assessed by flow cytometry. BMDC were incubated with labeled proteins on ice before being analyzed by flow cytometry. BMDC not incubated with proteins are depicted as a grey filled.

Image from Del Cid N et al., PLoS One, 7(7):e41727. Fig 2.; doi: 10.1371/journal.pone.0041727. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - APC Conjugation Kit - Lightning-Link® (AB201807)

APC Conjugation Kit - Lightning-Link® labeling anti-human 4β7 antibody for Flow cytometry.

McLinden RJ et al. used ab201807 as part of examining HIV-1 neutralizing antibodies.

They used the kit to conjugate APC to anti-human 4β7 antibody for use in flow cytometry.

A. Flow cytometric analysis of CD4, CCR5 and α4β7 expression in the A3R5.7 cell line. 0.5 x 106 cells were singly stained for 30 minutes with fluorochrome-conjugated antibodies as shown followed by fixation in 2% paraformaldehyde. Data are representative of at least two independent experiments. Isotype controls are shown in grey. Nearly all cells were positive for CD4 and CCR5 while approximately half were positive for α4β7.

B. Comparison of cell surface CD4, CCR5 and α4β7 receptor densities in various cell targets. 0.5 x 106 cells were stained with fluorochrome-conjugated antibodies and compared to defined populations of similarly stained Quantum Simply Cellular beads. PBMC were stimulated with CD3.8 bi-specific antibody in the presence of 50U/mL rhIL-2. Assuming monovalent antibody-to-surface receptor binding, the Antibody Binding Capacity (ABC) calculated is equivalent to receptors/cell. Data represents the mean of two separate experiments. TZM-bl cells express high levels of CD4 and CCR5 but are negative for α4β7 while A3R5.7 cells possess CCR5 and α4β7 densities more similar to PBMC. CD4 expression on TZM-bl was beyond assay range.

Image from McLinden RJ et al., PLoS One, 8(11):e77756. Fig 2 .; doi: 10.1371/journal.pone.0077756. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

• Conjugation

PubMed

Conjugation - APC Conjugation Kit - Lightning-Link® (AB201807)

APC Conjugation Kit - Lightning-Link® labeling IL-25 and TNFR2 extracellular domain for Flow cytometry.

Starkie DO et al. used ab201807 to identify antigen-specific mouse memory B cells from TNFR2 and IL-25 immunised mice.

Image from Starkie DO et al., PLoS One, 11(3):e0152282. Fig 3 and 2.; doi: 10.1371/journal.pone.0152282. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/