Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-Beta Tubulin monoclonal antibody (Cat.NO. 66240-1-Ig, isotype IgG2a). Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection. Different devices were used for imaging for the same membrane.

Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-ZNF174 monoclonal antibody (Cat.NO. 68426-1-Ig, isotype IgG3). Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection. Different devices were used for imaging for the same membrane.

Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-EIF3E monoclonal antibody (Cat.NO. 67095-1-Ig, isotype IgG1) at dilution of 1:50000. Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection.

Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-Catalase monoclonal antibody (Cat.NO. 66765-1-Ig, isotype IgG1) at dilution of 1:10000. Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection.

Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-Beta Tubulin monoclonal antibody (Cat.NO. 66240-1-Ig, isotype IgG2a) at dilution of 1:100000. Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection.

Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-FAK monoclonal antibody (Cat.NO. 66258-1-Ig, isotype IgG2c) at dilution of 1:5000. Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection.

Various lysates were subjected to SDS-PAGE followed by western blot with mouse anti-ZNF174 monoclonal antibody (Cat.NO. 68426-1-Ig, isotype IgG3) at dilution of 1:2000. Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:10000 for detection.

Immunofluorescence of A431 cells: PFA-fixed cells were stained with anti-B23 (60096-1-Ig), followed by Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM006, 1:500, red), nuclei stained with DAPI (cyan). Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich. Device setting for target: Laser Ex max 764nm, filter Em max 775nm.

Immunofluorescence of A431 cells: PFA-fixed cells were co-stained with mouse anti-B23 (60096-1-Ig) and rabbit anti-Lamin B1 (12987-1-AP), followed by Multi-rAb CoraLite® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM006, 1:500, red) and Nano-Secondary srb2GCL555-1 (yellow). Confocal images were acquired with a 63x oil objective and post-processed. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich. Device setting for RGAM006: Laser Ex max 764nm, filter Em max 775nm.

Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11 ) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 750-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (RGAM006), CL488-Phalloidin (green).

1X10^6 MOLT4 were surface stained with 0.2 ug Anti-Human CD8 (65204-1-Ig, Clone: UCHT4) and Mouse IgG2a Isotype Control 66360-2-Ig. Multi-rAb CoraLite® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) RGAM006 was used at 1:500 for detection.

Live A431 cells were incubated with anti-human EGFR (Cetuximab biosimilar) followed by Nano-Secondary® alpaca anti-human IgG, recombinant VHH, CoraLite® Plus 750 [CTK0117] (shuGCL750-2, gray). Cells were fixed and nuclei were stained with DAPI (blue). Epifluorescence images were acquired with a 40x objective and post-processed.

Immunofluorescence analysis of transgenic HeLa cells stably expressing H2B-myc. PFA-fixed cells were stained with anti-c-myc tag antibody and Nano-Secondary® alpaca anti-human IgG, recombinant VHH, CoraLite® Plus 750 [CTK0117] (shuGCL750-2, magenta).​

Schematic representation of anti-human IgG Nano-Secondary binding to its target. Human IgG primary antibody (blue) is bound with two identical monoclonal human IgG Fc- specific Nanobody molecules (green). In total, 4 fluorophores (red stars) label the human IgG primary antibody.

Immunofluorescence analysis of HeLa cells stained with mouse IgG1 anti-B23 antibody and Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, CoraLite® Plus 750 (smsG1CL750-1, white). Nuclei were stained with DAPI (blue).

HEK-293 cell lysates were subjected to SDS-PAGE followed by multiplex western blot analysis with 3 mouse primary antibodies including anti-ATP50 (66696-1-Ig), anti-AIF (67791-1-Ig), and anti-actin (66009-1-Ig). Primary antibodies were detected using 3 mouse IgG subclass-specific nano-secondary reagents including Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, CoraLite® Plus 750 (smsG1CL750-1, white), Nano-Secondary® alpaca anti-mouse IgG2a, recombinant VHH, CoraLite® Plus 488 (smsG2aCL488-1, green), and Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, CoraLite® Plus 555 (smsG2bCL555-1, red).

Schematic representation of anti-mouse IgG2a Nano-Secondary binding to its target. Mouse IgG2a primary antibody (grey) is bound with two identical monoclonal mouse IgG2a Fc- specific Nanobody molecules (green). In total, 4 fluorophores (red stars) label the mouse IgG2a primary antibody.

The alpaca anti-mouse IgG2a Fc-specific recombinant VHH [CTK0114] has a very high affinity (KD <10 pM) for mouse IgG2a primary antibody.

Immunofluorescence analysis of Hela cells stained with mouse IgG2a anti-Lamin B2 antibody (66240-1-Ig) and Nano-Secondary® alpaca anti-mouse IgG2a, recombinant VHH, CoraLite® Plus 750 (magenta). Nuclei were stained with DAPI (blue). Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.

Flow cytometry analysis of 1X10^6 human peripheral blood mononuclear cells (PBMCs) stained with anti-human CD3 (clone OKT3, 65133-1-Ig) and Nano-Secondary® alpaca anti-mouse IgG2a, recombinant VHH, CoraLite® Plus 750.

Immunofluorescence analysis of HeLa cells stained with mouse IgG2b anti-IMP3 antibody (66247-1-Ig) and Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, CoraLite® Plus 750 (smsG1CL750-1, gray). Nuclei were stained with DAPI (blue). 

HEK-293 cell lysates were subjected to SDS-PAGE followed by multiplex western blot analysis with 3 mouse primary antibodies including anti-TDP43 (60019-1-Ig), anti-tubulin (66240-1-Ig), and anti-IMP3 (66247-1-Ig). Primary antibodies were detected using 3 mouse IgG subclass-specific nano-secondary reagents including Nano-Secondary® alpaca anti-mouse IgG1, recombinant VHH, CoraLite® Plus 555 (smsG1CL555-1, red), Nano-Secondary® alpaca anti-mouse IgG2a, recombinant VHH, CoraLite® Plus 647 (smsG2aCL647-1, blue), and Nano-Secondary® alpaca anti-mouse IgG2b, recombinant VHH, CoraLite® Plus 750 (smsG2bCL750-1, gray).

HEK-293 and Hela cell lysates were subjected to SDS-PAGE followed by fluorescent western blot analysis with rabbit anti-GAPDH antiboy (10494-1-AP) and Nano-Secondary® alpaca anti-rabbit IgG, recombinant VHH, CoraLite® Plus 750 (srb2GCL750-1).

Immunofluorescence analysis of A431 cells stained with rabbit anti-tubulin antibody (80713-1-AP, cyan) and Nano-Secondary® alpaca anti-rabbit IgG, recombinant VHH, CoraLite® Plus 750 (srb2GCL750-1), 1:500. Nuclei were stained with DAPI. Images were recorded at the Core Facility Bioimaging at the Biomedical Center, LMU Munich.