Immunocytochemistry - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Direct immunofluorescence - Lightning-Link® Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610].

Direct immunofluorescent staining using Recombinant Anti-Ki67 antibody [EPR3610] - BSA and Azide free (ab209897) labelled with Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553).

Lightning-Link® Alexa Fluor® 488 Anti-Ki67 antibody [EPR3610] conjugate was used to stain Ki67-wild-type HAP1 cells (top panel) and Ki67-knock-out HAP1 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were incubated with 1 μg/ml of LL-Anti-Ki67 Alexa Fluor® 488 (shown in green) or ab209897 (Anti-Ki67 antibody, unlabelled control) overnight at +4°C. ab209897 treated cells only were incubated with ab150087 at 1/1000 dilution for 1 hour at room temperature (shown in red).

Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Immunocytochemistry - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Direct immunofluorescence - Lightning-Link® Alexa Fluor® 488 Anti-Vimentin antibody [EPR3776].

Direct immunofluorescent staining using Recombinant Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555) labelled with Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553).

Lightning-Link® Alexa Fluor® 488 Anti-Vimentin antibody [EPR3776] conjugate was used to stain Vimentin-wild-type HAP1 cells (top panel) and Vimentin-knock-out HAP1cells (bottom panel). The cells were fixed with 100% methanol at -20°C (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were incubated with 1 μg/ml of LL-Anti-Vimentin Alexa Fluor® 488 (shown in green) or ab193555 (Anti-Vimentin antibody, unlabelled control) overnight at +4°C. ab193555 treated cells only were incubated with ab150087 at 1/1000 dilution for 1 hour at room temperature (shown in red).

Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Immunocytochemistry - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Direct immunofluorescence - Lightning-Link® Alexa Fluor® 488 Anti-EGFR antibody [E235].

Direct immunofluorescent staining using Recombinant Anti-EGFR antibody [E235] - BSA and Azide free (ab227459) labelled with Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553).

Lightning-Link® Alexa Fluor® 488 Anti-EGFR antibody [E235] conjugate was used to stain wild-type A431 cells (top panel) and MCF7 negative cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) or 100% methanol at -20°C (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h.

The cells were incubated with 1 μg/ml of LL-Anti-EGFR Alexa Fluor® 488 conjugate (shown in green) or ab227459 (Anti-EGFR antibody, unlabelled control) overnight at +4°C. ab227459 treated cells only were incubated with ab150087 at 1/1000 dilution for 1 hour at room temperature (shown in red).

Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Immunocytochemistry - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Direct immunofluorescence - Lightning-Link® Alexa Fluor® 488 Anti-Cytokeratin 19 antibody [EP1580Y].

Direct immunofluorescent staining using Recombinant Anti-Cytokeratin 19 antibody [EP1580Y] - BSA and Azide free (ab195872) labelled with Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553).

Lightning-Link® Alexa Fluor® 488 Anti-Cytokeratin 19 [EP1580Y] conjugate was used to stain wild-type MCF7 cells (top panel) and A431 negative cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) or 100% methanol at -20°C (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were incubated with 1 μg/ml of LL-Anti-Cytokeratin 19 Alexa Fluor® 488 (shown in green) or ab195872 (Anti-Cytokeratin 19 antibody, unlabelled control) overnight at +4°C. ab195872 treated cells only were incubated with ab150087 at 1/1000 dilution for 1 hour at room temperature (shown in red).

Nuclear DNA was labelled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Flow Cytometry - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Alexa Fluor® 488 Conjugation Kit (Fast).

Bardhan, Kankana, et al used Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553) as part of examining PD-1pY248+ (pPD-1) expression in human T cells. They used the kit to conjugate Alexa Fluor® 488 to Anti-PD-1pY248 antibody for use in flow cytometry.

pPD-1 is predominantly expressed in CD8+ TCM cells. CCR7 and CD45RO markers were used to identify central memory (TCM) and effector memory (TEM) T cells. After gating on CD45RO expression, TCM (CD45RO+CCR7+) and TEM (CD45RO+CCR7−) CD4+ and CD8+ T cells were identified by assessing expression of CCR7. In TCM and TEM populations, expression of PD-1 was determined and, subsequently, expression of pPD-1 (pPD-1-Y248) was assessed in the PD-1+ population within each subset. Results are representative of six separate experiments.

Image from Bardhan, Kankana, et al., Scientific reports, 9(1):17252. doi: 10.1038/s41598-019-53463-0. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

Immunohistochemistry (Frozen sections) - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Alexa Fluor 488 Conjugation Kit.

Soldevila, Ferran et al used Alexa Fluor® 488 Conjugation Kit - Lightning-Link® (ab236553) as part of examining myeloid cells in the porcine palatine tonsil. They used the kit to conjugate Alexa Fluor® 488 to anti-pig CD4α antibody for use in confocal microscopy.

In situ localization of the CD14+ cells and plasmocytoid dendritic cells in porcine palatine tonsil. CD14+ cells and pDCs were localized by confocal microscopy following ethanol fixation of tonsil slices. The areas assessed included the follicle (F), the interfollicular region (IFA), the crypt (C). The tissue was stained using panel 1 antibodies; white arrow CD14+ cells and yellow arrow pDCs. Images are representative of at least two images from each section, from three different pigs. Objective used: (A) 63× oil immersion. Scale bars as shown.

Image from Soldevila, Ferran et al. Frontiers in immunology vol. 9 1800. 15 Aug. 2018, doi:10.3389/fimmu.2018.01800. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

Conjugation - Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553)

Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link®.

A polyclonal antibody was purchased from a commercial source and conjugated to Alexa Fluor® 488 using Alexa Fluor® 488 Conjugation Kit (Fast) - Lightning-Link® (ab236553) and to CF™488A dye using a competitor conjugation kit. The conjugates were tested by ELISA and the fluorescence measured at 523nm after excitation at 487nm. The Abcam conjugate out-performs the competitor.