Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed

41 product images

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-beta III Tubulin antibody [TU-20] - Neuronal Marker ab7751 staining beta III Tubulin in Neuro-2a cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Anti-beta III Tubulin antibody [TU-20] - Neuronal Marker ab7751 at 1/1000 and Anti-beta Tubulin antibody - Loading Control ab6046 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Mouse secondary (ab150117) at 2 μg/ml (shown in green) and AlexaFluor®594 Goat anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
  Negative controls: 1, Rabbit primary and anti-mouse secondary antibody; 2 , Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-GAPDH antibody [6C5] - Loading Control ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

  The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with Anti-GAPDH antibody [6C5] - Loading Control ab8245 at 5 μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046 at 1 μg/ml overnight at +4°C followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.

  Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha-Tubulin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at a working concentration of 0.5μg/ml and Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
  This product also gave a positive signal in 100% methanol (5 min) fixed SV40 cells under the same testing conditions.
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha-Tubulin in NIH3T3 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μl/ml and Anti-beta Tubulin antibody - Loading Control ab6046 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green) and anti-rabbit AlexaFluor® 594 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
  Negative controls: 1, Rabbit primary antibody and anti-mouse secondary antibody; 2 , Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-GAPDH antibody [6C5] - Loading Control ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.

  The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with Anti-GAPDH antibody [6C5] - Loading Control ab8245 at 1 μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272 at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

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

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 staining alpha-Tubulin in Caco-2 cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μg/ml and Anti-beta Tubulin antibody - Loading Control ab6046 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green) and anti-rabbit AlexaFluor® 594 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
  Negative controls: 1, Rabbit primary antibody and anti-mouse secondary antibody; 2 , Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-GAPDH antibody [6C5] - Loading Control ab8245 staining GAPDH in SV40LT-SMC (Rat SV40-transfected aorta smooth cell line) cells.

  The cells were fixed with 4% formaldehyde (10 minutes) permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with Anti-GAPDH antibody [6C5] - Loading Control ab8245 at 5μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272 at 1/250 dilution overnight at +4°C followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.

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

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  ICC/IF image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 10μg/ml) overnight at +4°C. The secondary antibody (green) was ab150117 Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at 1μg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.

  The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.

• 

  This image is courtesy of Dr. Shaohua Li

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School

  Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)

  Preparation:

  Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 μm thickness

  Primary antibody 1: Mouse anti-Ki67 (Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker ab53280), 1:50

  Primary antibody 2: Rabbit anti-laminin, 1:400
  Secondary antibody 1: Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150117), 1:200

  Secondary antibody 2: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) pre-adsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150084), 1:300
  Nuclei were counterstained with DAPI

• 

  Alexa Fluor® - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)
• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling Hsp60 with Anti-Hsp60 antibody [24/HSP60] - Mitochondrial Marker ab300659 at 1/250 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mitochondrial staining in RAW 264.7 cell line is observed. Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker ab186735 Anti-TOMM20 Rabbit monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse splenocyte cells labelling MHC Class II with Anti-MHC Class II antibody [MRC OX-6] ab23990 at 1/100 dilution (10.63 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used at 1/100 dilution (5µg/mL) as counterstain for tubulin (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody.
  
  Confocal image showing membranous and cytoplasmic staining in subsets of mouse splenocyte . Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling CD40L with Anti-CD40L antibody [AT161-10] ab281905 at 1/100 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/200 dilution.

  Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor^® 594) was used as a counterstain at a 1/1000 dilution.

  Confocal image showing membranous staining in subsets of human PBMCs treated with Phorbol-12-myristate-13-acetate (50 ng/mL) and Ionomycin (500 ng/mL) for 6 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat splenocyte cells labelling MHC Class II with Anti-MHC Class II antibody [MRC OX-6] ab23990 at 1/100 dilution (10.63 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used at 1/100 dilution (5µg/mL) as counterstain for tubulin (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody.
  
  Confocal image showing membranous and cytoplasmic staining in subsets of rat splenocyte.
  Negative control: C6.
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  NIH/3T3 cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1%Triton X-100 permeabilized HDLM-2 (human Hodgkin lymphoma cell) cells labelling Semaphorin 4D/CD100 with Anti-Semaphorin 4D/CD100 antibody [30/CD100] ab307685 at 1/100 dilution (9.97 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green).
  
  Confocal image showing membranous staining in HDLM-2 cell line, no staining was observed in PC-3 cell line.
  
  Negative control: PC-3 (PMID: 23775445).
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue).
  
  Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized C6 (rat glial tumor glial cell) cells labelling Hsp60 with Anti-Hsp60 antibody [24/HSP60] - Mitochondrial Marker ab300659 at 1/250 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mitochondrial staining in C6 cell line is observed. Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker ab186735 Anti-TOMM20 Rabbit monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 10 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1%Triton X-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling P cadherin with Anti-P cadherin antibody [56/P-Cadherin] ab307740 at 1/100 dilution (8.94 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green).
  
  Confocal image showing membranous staining in A431 cell line, no staining was observed in MCF7 cell line.
  
  Negative control: MCF7 (PMID:10545506).
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue).
  
  Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SIRT1 KO A549 (SIRT1 knockout human lung carcinoma epithelial cell) cells labelling SIRT1 with Anti-SIRT1 antibody [19A7AB4] ab110304 at 1/100 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

  Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor^® 594) was used as a counterstain at a 1/200 dilution.

  Confocal image showing nuclear staining in parental A549 cell line and no staining in SIRT1 KO A549 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells transfected with a HIV1 Rev M, HIV Rev N, a HIV1 Rev O and HIV2 Rev expression vector labelling HIV Rev with Anti-HIV1 Rev antibody [Rev-6] ab85529 at 1/500 dilution. Anti-HIV1 Rev antibody [Rev-6] ab85529 was incubated overnight at 4° C, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 1/1000 dilution at RT for 45 min. Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody at 1/200 dilution, was co-incubated with Anti-HIV1 Rev antibody [Rev-6] ab85529 overnight at 4° C. Nucleus were visualized using DAPI.

  Confocal image showing positive staining in 293T cells transfected with a HIV1 Rev M and HIV Rev N expression vector containing a myc-His-tag®(shown in green). but no staining in 293T cells transfected with a HIV1 Rev O and HIV2 Rev expression vector containing a myc-His-tag®.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1%Triton X-100 permeabilized Saos-2 (human osteosarcoma epithelial cell) cells labelling NPAT with Anti-NPAT antibody [27/NPAT] ab307837 at 1/250 dilution (3.99 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green).
  
  Confocal image showing nuclear foci staining in Saos-2 cell line.
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 dilution (5 ug/ml) (Red). The nuclear counterstain was DAPI (Blue).
  
  Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblast) cells labelling SIRT1 with Anti-SIRT1 antibody [19A7AB4] ab110304 at 1/100 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

  Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor^® 594) was used as a counterstain at a 1/200 dilution.

  Confocal image showing nuclear staining in C2C12 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling SIRT1 with Anti-SIRT1 antibody [19A7AB4] ab110304 at 1/100 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

  Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor^® 594) was used as a counterstain at a 1/200 dilution.

  Confocal image showing nuclear staining in PC-12 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling Hsp60 with Anti-Hsp60 antibody [24/HSP60] - Mitochondrial Marker ab300659 at 1/250 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mitochondrial staining in HeLa cell line is observed. Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker ab186735 Anti-TOMM20 Rabbit monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MOLT-4 cells labelling PD1 with Anti-PD1 antibody [NAT105] ab52587 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed (ab150117) at 1/1000 dilution at RT for 45 min. Recombinant Alexa Fluor^® 594 Anti-beta Tubulin antibody [EPR16774] (Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369) was used as a counterstain at 1/50 dilution and was co-incubated with Anti-PD1 antibody [NAT105] ab52587 overnight at 4° C. Nucleus were visualized using DAPI. Confocal image showing membranous staining in MOLT-4 cells treated with 500 ng/ml Ionomycin and 10 ng/ml Phorbol-12-myristate-13-acetate (PMA) for 24 hours, and no staining in MOLT-4 cells.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-HIF-1 alpha antibody [H1alpha67] ab1 staining HIF-1 alpha in HeLa DFO cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-HIF-1 alpha antibody [H1alpha67] ab1 at 10µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 4% paraformaldehyde (10 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-HIF-1 alpha antibody [ESEE122] ab8366 staining HIF-1 alpha in Hela cells. Untreated and BrdU treated (10µM for 24 hours) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-HIF-1 alpha antibody [ESEE122] ab8366 at 10µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
  

  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling Cytokeration 19 with Anti-Cytokeratin 19 antibody [EP1580Y] - Mouse IgG2a (Chimeric) ab323562 at 1/800 (1.096 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing cytoplasmic staining in MCF7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
  
  Negative control: SH-SY5Y.
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab202272) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• 

  Immunohistochemistry (Frozen sections) - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescence staining of GLP1R in sections of frozen normal rat pancreas.

  Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with Anti-GLP-1R antibody [EPR21819] - Mouse IgG2a (Chimeric) ab324027 at 1/100 dilution and then incubated for 1 hour with ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

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

  For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescence staining of GLP1R in sections of formalin-fixed paraffin-embedded normal rat pancreas.

  Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with Anti-GLP-1R antibody [EPR21819] - Mouse IgG2a (Chimeric) ab324027 at 1/100 dilution and then incubated for 1 hour with ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

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

  For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-SHANK3 antibody [N69/46] ab193307 staining SHANK3 in Rat Primary Neurons DIV14 cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal rabbit serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-SHANK3 antibody [N69/46] ab193307 at 1µg/ml (shown in green) and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control (shown in pseudocolour magenta). Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

  Also suitable in cells fixed with 100% methanol (5 min).

  Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized transfected 293T (human embryonic kidney epithelial cell) cells labelling HIV Vif with Anti-HIV1 Vif antibody [319] ab66643 at 1/100 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluo^® 488) at 1/1000 dilution (green).

  Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) preadsorbed at a 1/1000 dilution (Magenta).

  Confocal image showing positive staining in 293T cells transfected with human immunodeficiency virus type 1 group M subtype B (isolate YU-2) (HIV-1) Vif expression vector containing a myc-His-tag^® (shown in green) but no staining in 293T cells transfected with a human immunodeficiency virus type 1 group M subtype K (isolate 96CM-MP535) (HIV-1) Vif, human immunodeficiency virus type 1 group M subtype K (isolate 97ZR-EQTB11) (HIV-1) Vif, human immunodeficiency virus type 1 group M subtype D (isolate ELI) (HIV-1) Vif expression vector containing a myc-His-tag^®. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
  A: 293T cells transfected with an empty vector containing a myc-His-tag^®
  B: 293T cells transfected with a human immunodeficiency virus type 1 group M subtype K (isolate 96CM-MP535) (HIV-1) Vif expression vector containing a myc-His-tag^®
  C: 293T cells transfected with a human immunodeficiency virus type 1 group M subtype K (isolate 97ZR-EQTB11) (HIV-1) Vif expression vector containing a myc-His-tag^®
  D: 293T cells transfected with a human immunodeficiency virus type 1 group M subtype B (isolate YU-2) (HIV-1) Vif expression vector containing a myc-His-tag^®
  E: 293T cells transfected with a human immunodeficiency virus type 1 group M subtype D (isolate ELI) (HIV-1) Vif expression vector containing a myc-His-tag^®

• 

  Flow Cytometry (Intracellular) - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Flow cytometry overlay histogram showing mutant p53 in A-431 cells stained with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) (1x 106 cells in 100μl at 0.04μg/ml (1/25000)) for 30min at 22°C. The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) was incubated at 1/4000 for 30min at 22°C. Isotype control antibody (black line) Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody gave a positive signal in A-431 fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Anti-alpha smooth muscle Actin antibody [1A4] ab7817 staining alpha smooth muscle Actin in wild-type HeLa cells (top panel) and ACTA2 knockout HeLa cells (Human ACTA2 knockout HeLa cell line ab264014) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-alpha smooth muscle Actin antibody [1A4] ab7817 at 5ug/ml concentration and Anti-beta Tubulin antibody - Loading Control ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 ug/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 2 ug/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

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

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1%Triton X-100 permeabilized A431 (human epidermoid carcinoma epithelial cell) cells labelling NPAT with Anti-NPAT antibody [27/NPAT] ab307837 at 1/250 dilution (3.99 ug/ml), followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2ug/ml) (Green).
  
  Confocal image showing nuclear foci staining in A431 cell line.
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] - Microtubule Marker ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (10ug/ml) (Red). The nuclear counterstain was DAPI (Blue).
  
  Secondary antibody only control: Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2ug/ml).

• 

  Flow Cytometry (Intracellular) - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Flow cytometry overlay histogram showing wild-type p53 in Hek-293 positive cells (left) and MCF7 negative cells (right) stained with Anti-p53 antibody [DO-1] - ChIP Grade ab1101 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) (1x 106 cells in 100μl at 0.2μg/ml (1/5000)) for 30min at 22°C. The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) was incubated at 1/4000 for 30min at 22°C. Isotype control antibody (black line) Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) was used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody gave a positive signal in Hek-293 fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  HeLa cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Human papillomavirus 6 L1 protein with Anti-Human papillomavirus 6 L1 protein antibody [LH6L1_D] ab322925 at 1/2000 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing nuclear staining in 293T cells transfected with a human papillomavirus type 6B L1, (shown in green) but no staining in 293T cells transfected with a human papillomavirus type 11 L1 expression vector containing a myc tag. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
  

  Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Human papillomavirus 6 L1 protein with Anti-Human papillomavirus 6 L1 protein antibody [LH6L1_D] ab322925 at 1/2000 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing nuclear staining in 293T cells transfected with a human papillomavirus type 6B L1 expression vector containing a myc tag (shown in green) but no staining in 293T cells transfected with a human papillomavirus type 18 L1 or human papillomavirus type 4 L1 expression vector containing a myc tag. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling HPV4 L2 with Anti-Human papillomavirus type 4 L2 antibody [HPV4L2(20-38)_4SA] ab322958 at 1/100 dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing nuclear staining in 293T cells transfected with a Human papillomavirus type 4 L2 and Human papillomavirus type 65 L2 expression vector containing a myc-His-tag®(shown in green) but no staining in 293T cells transfected with a human papillomavirus type 60 L2 expression vector containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• 

  Flow Cytometry (Intracellular) - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

  Flow cytometry overlay histogram showing mutant p53 in wild-type HAP1 (green line) and TP53 knockout HAP1 cells (red line) stained with Anti-p53 antibody [DO-1] - ChIP Grade ab1101. The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [DO-1] - ChIP Grade ab1101) (1x 106 cells in 100μl at 0.04 μg/ml (1/25000)) for 30min at 22°C.The secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) was incubated at 1/4000 for 30min at 22°C. Isotype control antibody Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) was used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line, TP53 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This antibody gave a positive signal in HAP1 fixed with 80% methanol (5 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.