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

41 product images

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left panel) / Caco-2 (Human colorectal adenocarcinoma epithelial cell, Right panel) using Anti-CD133 antibody [RM1002] - Stem Cell Marker ab278053 at 1/500 dilution (0.1µg) (red). The isotype control used was the Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730), black line and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Secondary antibody used was the Goat anti-rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution.

  Negative control: HEK-293T. (PMID 20167130)
  Gated on viable cells.

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Overlay histogram showing Jurkat cells stained with Anti-CD3 epsilon antibody [SP7] ab16669 (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-CD3 epsilon antibody [SP7] ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (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 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  ICC/IF image of beta Tubulin staining in HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat 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 (green) was ab150077 Alexa Fluor^® 488 goat anti-rabbit 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.

  IHC - Wholemount - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  IHC - Wholemount of Caenorhabditis elegans larvae labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody ab5095. The sample was incubated with primary antibody (1/500 in PBS + 3% BSA + 0.1% Triton X-100) for 12 hours at 4°C. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/1000), was used as the secondary antibody.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry/ Immunofluorescence analysis of C6(Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] ab32575 at 1/100 dilution (0.71 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunohistochemistry (Frozen sections) analysis of rat stomach tissue (4% PFA, 0.2% Tritron x-100) labelling Integrin alpha 6 with Anti-Integrin alpha 6 antibody [EPR18124] - BSA and Azide free ab240252 at 1/100 dilution. ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) was used as the secondary antibody at 1/1000 dilution. Negative control using PBS instead of primary antibody (left). Counterstained with DAPI.
  

  Positive staining was seen on rat stomach.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry analysis of SK-OV-3 (human ovarian cancer epithelial cell)cells labelling Zic2 with Anti-Zic2 antibody [EPR7790] ab150404 at 2.5 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Overlay histogram showing HeLa cells stained with Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade ab32356 (red line). The cells were fixed with 80% methanol (5 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-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade ab32356, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10^6 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 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Overlay histogram showing MCF7 cells stained with unpurified Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade ab32063 (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-Estrogen Receptor alpha antibody [E115] - ChIP Grade ab32063, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3(Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] ab32575 at 1/500 dilution (5.2 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Estrogen Receptor alpha with purified Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade ab32063 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) using Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei counterstained with DAPI (blue).
  Control 1: primary antibody (1/1000) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
  Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry analysis of Jurkat (human T cell leukemia T lymphocyte) labeling p95/NBS1 with purified Anti-p95/NBS1 (phospho S343) antibody [EP178] ab109453 at 1/100 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/1000 (2 μg/ml) was used as the secondary antibody. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.27 μg/ml) was used as counterstain. Nuclei were stained blue with DAPI. Negative control: PBS instead of the primary antibody.

  Confocal image showing increased nuclear staining in Jurkat cells treated with Etoposide (25uM) for 2 h.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 25 ug/mL anisomycin for 30 min, then Lambda Protein Phosphatase 31 for 2 hours cells labeling Hsp27 with purified Anti-Hsp27 (phospho S78) antibody [Y175] ab32501 at 1/50 dilution (11.3 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/mL) dilution. Goat anti-rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling KDM3B / JMJD1B with Anti-KDM3B / JMJD1B antibody [EPR24219-30] ab271046 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubμue Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry/Immunofluorescence analysis of Raji (human Burkitt's lymphoma) cells labelling HLA A with purified Anti-HLA A antibody [EP1395Y] ab52922 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
  Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
  Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling LAMP1 with Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (green). Confocal image showing cytoplasmic staining in HeLa. Anti-LAMP1 antibody [H4A3] - Lysosome Marker ab25630 Anti-LAMP1 mouse monoclonal antibody was used to counterstain at 1/500 (red). The Nuclear counterstain was DAPI (blue).

  Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with Purified Anti-ATG9A antibody [EPR2450(2)] ab108338 at 1/100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labeling KDM3B / JMJD1B with Anti-KDM3B / JMJD1B antibody [EPR24219-30] ab271046 at 1/500 dilution, followed by 150077 Goat Anti-Rabbit IgG H&L (Alexa Fluo^r® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
  

  Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labeling alpha smooth muscle Actin (green) with purified Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] ab32575 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Alexa Fluor®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).
  For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) were used. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  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, 1μg/ml) and (Anti-Lamin B1 antibody - Nuclear Envelope Marker ab16048, 1μg/ml) overnight at +4°C. The secondary antibodies were Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) ab150115 Alexa Fluor® 647 (red) goat anti-mouse IgG (H+L) used at 2μg/ml for 1h and ab150077 Alexa Fluor® 488 (green) goat anti-rabbit IgG (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei.

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  Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
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  ELISA - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  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® 488) (ab150077)

  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® 488) (ab150077)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LAMP1 KO HAP1 cells labelling LAMP1 with Anti-LAMP1 antibody [EPR24395-31] - Lysosome Marker ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 2 ug/ml dilution (green). Confocal image showing cytoplasmic staining in parental HAP1 cell line and no staining in LAMP1 KO HAP1 cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody- Microtubule Marker (Alexa Fluor ^® 594) was used to counterstain at 1/200 (red).

  Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblasts myoblast) cells labelling CD63 with Anti-CD63 antibody [RM1095] ab315108 at 1/100 (5.24 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (2 ug/ml) dilution (Green).

  Confocal image showing cytoplasmic staining in C2C12 cells. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

  Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/ml) dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) labeling BMAL1 with Anti-BMAL1 antibody [EPR23696-22] ab230822 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) (ab150077) secondary antibody at 1/1000 dilution (green).

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 was used as a counterstain.

  The nuclear counterstain is DAPI (blue).

  Confocal image showing positive staining in PC-3 cell line.

  Negative Control: Raji (PMID: 29230015).

  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® 488) (ab150077)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (Human colorectal adenocarcinoma epithelial cell) labeling 15-PGDH with Anti-15-PGDH antibody [EPR14332] ab187160 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 was used as a counterstain (red).

  The nuclear counterstain is DAPI (blue).

  Confocal image showing mainly cytoplasmic staining in SW480 cell line.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF-7 (human breast adenocarcinoma epithelial cell) labeling 15-PGDH with Anti-15-PGDH antibody [EPR14332] ab187160 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 was used as a counterstain (red).

  The nuclear counterstain is DAPI (blue).

  Confocal image showing mainly cytoplasmic staining in MCF-7 cell line.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) labelling ROR beta/RORB with Anti-ROR beta/RORB antibody [EPR15552] - N-terminal ab187657 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488 (ab150077) secondary antibody at 1/1000 dilution (green).

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution was used as a counterstain (red).

  The nuclear counterstain is DAPI (blue).

  Confocal image showing nuclear staining in HepG2 cell line. 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® 488) (ab150077)

  Immunofluorescent analysis was performed on 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse hippocampal primary neural/glia cells labelling Calbindin with Anti-Calbindin antibody [EP3478] ab108404 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody (ab150077) at dilution of 1/1000 (green).

  Anti-MAP2 antibody [HM-2] (Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267) at 1/1000 dilution was used as a counterstain, with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed as the secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at dilution of 1/1000 (red).

  Negative control 1: Anti-Calbindin (Anti-Calbindin antibody [EP3478] ab108404) at 1/250 dilution with Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed as the secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at dilution of 1/1000.

  Negative control 2: Anti-MAP2 antibody [HM-2] (Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267) at 1/200 dilution with Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody (ab150077) at dilution of 1/1000.

  The nuclear counterstain is DAPI (blue).

  Confocal image showing positive staining in subsets of mouse hippocampal primary neuron.

  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).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MDA-MB-231 (human breast adenocarcinoma epithelial cell) labeling FABP5 with Anti-FABP5 antibody [EPR22552-64] ab255276 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 was used as a counterstain (red).

  The nuclear counterstain is DAPI (blue).

  Confocal image showing cytoplasmic and nuclear staining in MDA-MB-231 cells.

  Negative Control: T-47D (PMID: 21356353).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 100% methanol fixed, 0.1% Triton X-100 permeabilized HUVEC (human umbilical vein endothelial) cells labelling IL-33 with Anti-IL-33 antibody [EPR26367-2] ab316846 at 1/500 dilution, followed by Goat anti-Rabbit (AlexaFluor^® 488) (ab150077) secondary antibody at 1/1000 dilution. Confocal image showing the nuclear and weak cytoplasmic staining (shown in green) is decreased after the treatment with IL-1β (5 ng/ml) for 24 hours in HUVEC cells. Nuclear DNA was labeled with DAPI (shown in blue).

  Counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A]-Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (shown in magenta).

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

  The reduction in IL-33 when stimulated with TNFa in HUVEC cells (specially the nuclear IL-33) (PMID: 18787100).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CLDN1 KO A431(CLDN1 knockout human epidermoid carcinoma epithelial cell) cells labelling Claudin 1 with Anti-Claudin 1 antibody [EPR25359-48] ab307692 at 1/250 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) antibody at 1/1000 dilution (Green).

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^®
  594) ( Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 ) was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is AlexaFluor488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

  Confocal image showing membranous and cytoplasmic staining in wildtype A431 cells and showing no staining in CLDN1 knockout A431 cells.
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling Irak3 with Anti-IRAKM antibody [EPR26376-117] ab307708 at 1/250 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) antibody at 1/1000 dilution.

  Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used to counterstain tubulin at 1/200 dilution. The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

• 

  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD2 with purified Anti-CD2 antibody [EPR6451] ab131276 at 1:20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• 

  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CSNK2A1 with purified Anti-CSNK2A1 antibody [EP1963Y] ab76040 at 1:20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• 

  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) transfected with HA tagged IL-21 construct cells labelling Il21 with Anti-IL-21 antibody [EPR22618-28] ab227837 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• 

  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Th17 polarized Mouse splenocytes were treated with 500ng/ml PMA and 500 ng/ml ionomycin for 1 hours, then add 1ug/ml Brefeldin A for another 4 hours cells labelling Il21 with Anti-IL-21 antibody [EPR22618-28] ab227837 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. IL21 is mutual exclusively expressed with IFN gamma (Th1 marker).Cells were provided by an anonymous collaborator.

• 

  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Intracellular Flow Cytometry analysis of 293T (human embryonic kidney epithelial cell) transfected with myc-tagged PADI4 construct, then treated with 10mM CaCl2 and 10uM Ionomycin for 4h labeling Histone H3 (citrulline R2 + R8 + R17) with Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody.
  Isotype control : Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)/left.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Intracellular Flow Cytometry analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling PSMA with Anti-PSMA antibody [EP3253] - BSA and Azide free ab247436 at 1/100 dilution (1 µg) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black). Unlabeled control - cell without incubation with primary antibody and secondary antibody (Blue).

• 

  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

  Flow cytometric analysis of P815 (mouse mastocytoma mast cell line, left) cells and C2C12 ( mouse myoblast, right) cells labeling CD80 with Anti-CD80 antibody [EPR22127-251] ab222702 at 1/5000 (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

  Gated on viable cells.

  Negative control: P815 (PMID: 9237108).