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

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

ICC/IF image of 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 (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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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

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 ab1101 at 0.2µg/ml and 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 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% TritonX-100 permeabilized HepG2 cells labelling CEACAM1 + CEACAM3 + CEACAM6 with ab104450 at 1/250 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (green).

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) used at a 1/1000 dilution (red).

Confocal image showing membranous staining in HepG2 cell line.
Negative control : HeLa (PMID : 11580753).

Nuclear DNA was labelled with DAPI (shown in blue).

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

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 ab12039 at 5µg/ml and 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 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)

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 ab1101 at 0.2µg/ml and 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 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)

MCF7/ HCT 116 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, ab70475 at 1 : 100 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Mouse secondary antibody (ab150113) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with 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)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 cells labelling EGFR with ab231 at 1/50 dilution followed by secondary antibody ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (green).

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) used at a 1/1000 dilution (red).

Confocal image showing strong membranous and weak cytoplasmic staining in A431 cells.
Low expression control : HEK-293 (PMID : 26368334).

Nuclear DNA was labelled with DAPI (shown in blue).

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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)

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 ab64165 at 1µg/ml and 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 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)

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 ab80522 at 1µg/ml and 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 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)

HeLa cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, 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. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with 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)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Hsp27 with ab2790 at 1/100 dilution, followed by AlexaFluor® 488-conjugated Goat anti-Mouse secondary antibody (ab150113) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with 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.

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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)

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 ab23336 at 5µg/ml and 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 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)

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 ab8366 at 10μg/ml and 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 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)

ab230171 staining Vimentin in HeLa-VIM 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 ab230171 at 1µg/ml and ab6046. Cells were then incubated with 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).

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)

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 ab31830 at 1µg/ml and 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 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)

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 ab177004 at 1µg/ml and 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 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)

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 ab1 at 10µg/ml and 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 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)

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 ab53931 at 5µg/ml and 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 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)

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 ab105459 at 5µg/ml and 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 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)

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 ab13575 at 1µg/ml and 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 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)

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 ab238673 at 5µg/ml and 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 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)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labelling SOX10 with ab218522 at 1/50 dilution followed by secondary antibody ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution (green).

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (red).

Confocal image showing nuclear staining in A375 cells.
Negative control : MCF7 (PMID : 23799842).
Nuclear DNA was labelled with DAPI (shown in 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 mouse primary neural/glia cells labelling Myelin Basic Protein with ab7349 at a 1/200 dilution, followed by Goat anti-Rat (Alexa Fluor^® 488) (ab150157) secondary antibody at a 1/1000 dilution. Confocal image showing positive staining in mouse primary oligodendroglia cells (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

Counterstained with Anti-MAP2 antibody (ab183830) at a 1/1000 dilution, followed by Goat Anti-Rabbit secondary (Alexa Fluor^® 594) (ab150080) at a 1/1000 dilution (shown in magenta).

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).

The negative controls are as follows :

-ve control 1 : Myelin Basic Protein (ab7349) at a 1/200 dilution, followed by Goat anti-Rabbit (Alexa Fluor^® 594) (ab150080) at a 1/1000 dilution.

-ve control 2 : Anti-MAP2 (ab183830) at a 1/1000 dilution, followed by Goat anti-Rat (Alexa Fluor^® 488) (ab150157) at a 1/1000 dilution.

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

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 ab78078 at 1µg/ml and 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 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% TritonX-100 permeabilized rat primary neurons labelling Synaptophysin with ab309493 at 1/100 dilution, followed by 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). 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 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)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neurons labelling Synaptophysin with ab309493 at 1/100 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in mouse 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). 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 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)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labelling Mouse IgG1 with ab280974 at 1/20 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). 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 : ab280974 at a 1/20 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2 : ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

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 ab53521 at 1µg/ml and ab6046. Cells were then incubated with 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)

RAW 264.7 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, ab53444 at 1 : 50 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Rat secondary antibody (ab150157) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with 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)

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 ab27591 at 10μg/ml concentration and 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 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)

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 ab90247 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with 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)

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.

• ICC/IF

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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, 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. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with 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.

• ICC/IF

Supplier Data

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, ab8878 at 1 : 50 was incubated overnight at 4° C, followed by AlexaFluor® 488- conjugated Goat anti-Rat secondary antibody (ab150157) at 1/1000 dilution at RT for 45 min. ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with 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)

ab53521 staining A2B5 in primary mouse neurons/glia, DIV14 (prepared from E18 mouse hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP) 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 ab53521 at 1µg/ml and ab6046. Cells were then incubated with 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

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 ab10321 at 1µg/ml and 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 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.

• ICC/IF

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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 mouse primary neuron cells labelling Glutamate Receptor 1 (AMPA subtype) with ab174785 at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 dilution at RT for 45 min. Anti-MAP2 antibody [EPR19691] (ab183830) was used as a counterstain at 1/1000 dilution and was co-incubated with ab174785 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.

• ICC/IF

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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 neuron cells labelling Glutamate Receptor 1 (AMPA subtype) with ab174785 at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 dilution at RT for 45 min. Anti-MAP2 antibody [EPR19691] (ab183830) was used as a counterstain at 1/1000 dilution and was co-incubated with ab174785 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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Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) (AB150080)

Immunofluorescence staining of Doublecortin in a section of frozen PFA perfusion fixed 8 week old mouse dentate gyrus.

After dissection, the tissue was further fixed in PFA overnight, washed in PBS and then incubated overnight in 30% sucrose. It was then embedded in OCT and cryosectioned. No antigen retrieval step was performed prior to staining. Performed on a Leica BOND^TM. The section was incubated at room temperature for 1 hour with ab18723 at 1ug/mL and then incubated with ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed, (Shown in magenta) 1/1000) for 1 hour at room temperature. 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.