Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594)

40 product images

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Overlay histogram showing Jurkat cells stained with Anti-CD45 antibody [EP322Y] ab40763 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-CD45 antibody [EP322Y] ab40763, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, 0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 561nm laser and 610/20 bandpass filter.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  ICC/IF image of Anti-beta Tubulin antibody - Loading Control ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the primary antibody (Anti-beta Tubulin antibody - Loading Control ab6046, 5μg/ml) overnight at +4°C. The secondary antibody (orange) was ab150080 Alexa Fluor^® 4594 goat anti-rabbir IgG (H+L) used at 2μg/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.

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  IHC-P image of alpha smooth muscle actin (Anti-alpha smooth muscle Actin antibody ab5694) staining E16.5 mouse embryo gut. Paraformaldehyde fixed and paraffin embedded E16.5 mouse embryo gut sections were dewaxed and rehydrated before antigen retrieval (4 mins in a pressure cooker in 10mM Tris/0.4mM EDTA buffer pH 9.5). They were then incubated in 50mM NH4Cl for 30 minutes and washed/blocked in 3x 10 minute washes of PBS containing 1% BSA + 0.2% gelatine and 0.05% saponin. Sections were incubated overnight with a primary antibody against alpha smooth muscle actin (Anti-alpha smooth muscle Actin antibody ab5694), diluted 1/250 in PBS containing 0.1% BSA and 0.3% triton. After 3 x 10 minute washes in of PBS containing 0.1% BSA, 0.2% gelatine and 0.05% saponin, the sections were incubated for 1 hr in the secondary antibody (ab150080, diluted 1/400, shown in red) and then the 3 washes repeated. Sections were mounted in Vectashield with DAPI (blue).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labelling Mouse IgG1 with Mouse IgG1 monoclonal [R312-MouseIgG1]-Isotype control ab280974 at 1/20 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).
  

  Confocal image showing no staining in RAW 264.7 cells.

  Negative control 1: Mouse IgG1 monoclonal [R312-MouseIgG1]-Isotype control ab280974 at a 1/20 dilution followed by ab150080 at a 1/1000 dilution.

  Negative control 2: Anti-beta Tubulin antibody [EPR16774] ab179513 at a 1/200 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 at a 1/1000 dilution.

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  Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
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  ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Cross-reactivity of the polyclonal secondary antibody Goat Anti-Rabbit IgG H&L ab182016 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 μg/ml (50 μl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Goat Anti-Rabbit IgG H&L ab182016 was then added starting at 1 μg/ml and gradually diluted 1/4 (50 μl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (Donkey Anti-Goat IgG H&L (HRP) ab6885) was used at 1/10,000 dilution (50 μl/well), followed by incubation for 1h at RT.
  

  For the batch tested, Goat Anti-Rabbit IgG H&L ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.

  This data was developed using the unconjugated antibody (Goat Anti-Rabbit IgG H&L ab182016).

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  ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Cross-reactivity of Goat anti-Rabbit IgG H&L (Goat Anti-Rabbit IgG H&L ab182016) and Goat anti-Rabbit IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 μg/ml (50 μl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 μg/ml and gradually diluted 1/4 (50 μl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (Donkey Anti-Goat IgG H&L (HRP) ab6885) was used at 1/10,000 dilution (50 μl/well), followed by incubation for 1h at RT. This data is from a representative dilution.
  

  This data was developed using the unconjugated antibody (Goat Anti-Rabbit IgG H&L ab182016).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-A2B5 antibody [105] ab53521 staining A2B5 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) 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 goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-A2B5 antibody [105] ab53521 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046. Cells were then incubated with Goat Anti-Mouse IgM mu chain (Alexa Fluor® 488) ab150121 at 1/1000 dilution (shown in green) and ab150080 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)
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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) transfected with GFP-tagged human HA Tag expression vector cells labelling HA tag with Anti-HA tag antibody [RM1058] ab314237 at 1/500 (1.042 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/1000 (2 ug/mL) dilution (Green).

  Confocal image showing cytoplasmic staining in 293T cells transfected with a human HA Tag expression vector containing a GFP tag.
  
  Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/mL) dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Cytokeratin 15 antibody [LHK15] ab80522 staining Cytokeratin 15 in A431 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-Cytokeratin 15 antibody [LHK15] ab80522 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Histone H2B antibody [mAbcam 64165] - ChIP Grade ab64165 staining Histone H2B in HeLa 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-Histone H2B antibody [mAbcam 64165] - ChIP Grade ab64165 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).

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

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-HOXB13 antibody [mAbcam53931] ab53931 staining HOXB13 in HepG2 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-HOXB13 antibody [mAbcam53931] ab53931 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).

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

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade ab31830 staining Histone H4 in HeLa 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-Histone H4 antibody [mAbcam 31830] - ChIP Grade ab31830 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-CD32B + CD32A antibody [AT10] ab23336 staining CD32B + CD32A in K-562 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 goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-CD32B + CD32A antibody [AT10] ab23336 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).

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

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Integrin alpha V+beta 5 antibody [P1F6] ab177004 staining Integrin alpha V+beta 5 in A549 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 goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-Integrin alpha V+beta 5 antibody [P1F6] ab177004 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).

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

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - BSA and Azide free ab238673 staining Histone H3 (phospho S10) in HeLa 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-Histone H3 (phospho S10) antibody [mAbcam 14955] - BSA and Azide free ab238673 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Cytochrome C antibody [7H8.2C12] ab13575 staining Cytochrome C in HepG2 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-Cytochrome C antibody [7H8.2C12] ab13575 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-MiTF antibody [C5] ab12039 staining MiTF in Malme-3M cells. The cells were 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-MiTF antibody [C5] ab12039 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).

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

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-WIPI2 antibody [2A2] ab105459 staining WIPI2 in HeLa Chloroquine treated 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-WIPI2 antibody [2A2] ab105459 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L Alexa Fluor^® 488) preadsorbed at 1/1000 dilution shown in green) and 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).

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

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  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 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] 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 (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  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 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] 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 (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  RAW 264.7 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-CD68 antibody [FA-11] ab53444 at 1:50 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Rat secondary antibody (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-CD68 antibody [FA-11] ab53444 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI. Confocal image showing cytoplasmic staining in RAW 264.7 cell line.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-DDX4 / MVH antibody [mAbcam27591] ab27591 staining DDX4/MVH in mES cells. 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-DDX4 / MVH antibody [mAbcam27591] ab27591 at 10μg/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) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Confocal image showing membranous staining in RAW 264.7 cells.
  
  RAW 264.7, WEHI-231 and THP-1 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-CD11b antibody [M1/70] ab8878 at 1:50 was incubated overnight at 4° C, followed by AlexaFluor® 488- conjugated Goat anti-Rat secondary antibody (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-CD11b antibody [M1/70] ab8878 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
  
  Negative control: WEHI-231 (PMID: 2457584)

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  MCF7/ HCT 116 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] ab70475 at 1:100 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Integrin beta 3 antibody [Y2/51] ab9509 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
  
  Confocal image showing membranous and cytoplasmic staining in MCF7 cells.
  Negative control: HCT 116 (PMID: 14998492)

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-Phosphotyrosine antibody [PY20] ab10321 staining Phosphotyrosine in C2C12 cells treated with 2mM H2O2 for 10mins. 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 goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-Phosphotyrosine antibody [PY20] ab10321 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).
  Also suitable in cells fixed with 100% methanol (5 min).
  Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-beta III Tubulin antibody [2G10] - Neuronal Marker ab78078 staining beta III Tubulin in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween 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-beta III Tubulin antibody [2G10] - Neuronal Marker ab78078 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).
  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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells labelling HLA DMB with primary antibody anti-HLA DMB (Anti-HLA DMB antibody [EPR7982] ab131273) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080) secondary antibody at 1/500 dilution. Confocal image showing cytoplasmic staining in 293T cells transfected with myc-tagged HLA-DMB expression vector, no staining in 293T cells transfected with myc-tagged NEK6 expression vector. Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 488) (Alexa Fluor® 488 Anti-Myc tag antibody [9E10] ab202008) was used as counterstain at 1/100 dilution. The nuclear counter stain is DAPI (blue).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 100% methanol permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Histone H3 (citrulline R2 + R8 + R17) with primary antibody anti-Histone H3 (citrulline R2 + R8 + R17) (Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584) at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080) secondary antibody at 1/1000 dilution. Confocal image showing nuclear staining on 293T cells transfected with a human PADI4 expression vector containing a GFP tag, then treated with 10 mM CaCl2 and 10 μM ionomycin for 2 hours. The nuclear counter stain is DAPI (blue).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Red. 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 goat serum/0.3M glycine in 0.1%PBS-Tween for 1h.
  Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining wild-type p53 in MCF7 cells (a low expressing cell line). 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-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).
  
  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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining wild-type p53 in Hek293 cells (a high expressing cell line). 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-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).
  
  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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-p53 antibody [DO-1] - ChIP Grade ab1101 staining mutant p53 in A431 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-p53 antibody [DO-1] - ChIP Grade ab1101 at 0.2µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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).
  
  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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Anti-F4/80 antibody [F4/80] - Macrophage Marker ab90247 staining F4/80 in Raw264.7 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween 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-F4/80 antibody [F4/80] - Macrophage Marker ab90247 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed ab150165, Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and 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 high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and 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.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neurons labelling Synaptophysin with Anti-Synaptophysin antibody [YE269] - Mouse IgG1 (Chimeric) ab309493 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green).
  
  Confocal image showing cytoplasmic staining in rat primary neuron cell line.
  
  Confocal scanning Z step was set as 0.3 µM followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Anti-MAP2 antibody [EPR19691] ab183830 Anti-MAP2 rabbit monoclonal antibody was used to counterstain tubulin at 1/1000 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/500 dilution (Red). The nuclear counterstain was DAPI (Blue).
  
  Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (ab150080)

  alpha smooth muscle Actin immunofluorescence staining of HeLa cells using mouse anti-alpha smooth muscle Actin antibody
  
  

  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) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 ug/ml (shown in green) and a goat secondary antibody to rabbit IgG (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).