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

41 product images

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometry overlay histogram showing wild-type HAP1 (green line) and TP53 knockout HAP1 cells stained with Anti-p53 antibody [SP161] ab227655 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [SP161] ab227655) (1x10⁶ in 100 µl at 0.008 µg/ml) for 30 min at 22°C.
  

  The secondary antibody Goat anti-Rabbit IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (ab150081) was used at 1/2000 for 30 min at 22°C.

  Isotype control antibody was Rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line TP53 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  This antibody gave a positive signal in TP53 HAP1 knockout cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  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, pre-adsorbed) (ab150081) was used at 1/2000 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) preadsorbed (ab150081)

  ICC/IF image of Anti-beta Actin antibody - Loading Control ab8227 stained HeLa cells. The cells were 4% formaldehyde fixed (10 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-beta Actin antibody - Loading Control ab8227, 5μg/ml) overnight at +4°C. The secondary antibody (green) was ab150081 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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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Anti-Ki67 antibody [EPR3610] ab92742 staining Ki67 in wild-type HAP1 cells (top panel) and Ki67 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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-Ki67 antibody [EPR3610] ab92742 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labeled in blue with DAPI.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-937 cells labelling IL-8 with Anti-IL-8 antibody [EPR26511-74] ab289967 at 1/100 (5.87 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 μg/mL) dilution (Green). Confocal image showing cytoplasmic staining is observed in U-937 cells treated with TPA (100 ng/mL) for 24 h, then LPS (5 μg/mL) for 7 h with Brefeldin A (300 ng/mL) for the last 3 h. 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 (2.5μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/mL) dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Unpurified Anti-Cyclin D1 antibody [EPR2241] - C-terminal ab134175 staining Cyclin D1 in MCF7 (Human breast adenocarcinoma cell line) cells treated with KN-93 (KN-93 (water soluble), CaMK II inhibitor ab120980).
  The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with Anti-Cyclin D1 antibody [EPR2241] - C-terminal ab134175 at 10μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Rabbit Alexa 488 secondary (ab150081) at 2 μg/ml (shown in green) and Goat anti-Mouse Alexa 594 secondary (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.
  Negative controls: 1- Rabbit primary and anti-mouse secondary antibody; 2 - Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis 4% Paraformaldehyde fixed 90% methanol permeabilised HEK-293T (Human embryonic kidney epithelial) cells transfected with a SARS-CoV-2 nsp2 (Left) or SARS-CoV2 nsp9 (Right) expression vector containing a myc tag labelling SARS-CoV-2 nsp9 with Anti-SARS-CoV-2 nsp9 antibody [EPR24855-25] ab284037 at 1/500 dilution. Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 80% Methanol fixed 0.1% TritonX-100 permeabilised U937 (Human histiocytic lymphoma monocyte) cells labelling CD36 with Anti-CD36 antibody [EPR22509-40] ab252922 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 1/1000 dilution. Confocal image showing membranous staining in U937 cells (Human histiocytic lymphoma monocyte) is observed. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) was used to counterstain Alpha Tubulin at 1/200 dilution.The Nuclear counterstain is DAPI .
  
  Secondary antibody only control: Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) at 1/1000 dilution.

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  This image is courtesy of a customer review submitted by Bryan Niedenberger

  Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Postnatal day 6 mouse testes were fixed with 4% paraformaldehyde. Tissue was embedded in O.C.T. and frozen. 5 micron sections were cut and transferred to slides. Sections were permeabilized with 0.1% Triton X-100 in PBS, and blocked with 3% BSA in 0.1% Triton X-100 + PBS. Sections were incubated with either (A) no primary antibody or (B ) anti-DDX4 (Anti-DDX4 / MVH antibody ab13840) for 1 h at RT. Sections were then washed 3X with 0.1% Triton X-100 in PBS and Goat-Anti Rabbit 488 (ab150081) applied at a 1/500 dilution. Sections were then mounted after washing 3X with 0.1% Triton X-100 in PBS.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-937 (Human histiocytic lymphoma monocyte) treated with 100ng/ml TPA for 24 hours, then 5μg/ml LPS for 4 hours, and add 300ng/ml BFA for another 3h (Red) / Untreated control (Green) cells labelling IL-8 with Anti-IL-8 antibody [EPR26511-74] ab289967 at 1/500 dilution (0.1μg) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

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  This image is courtesy of Dr. Shaohua Li

  Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

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

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

  Preparation:

  Fix in 3% PFA in PBS for 30 min at RT Incubate in 7.5% sucrose-PBS for 3h at RT Incubate in 15% sucrose-PBS at 4 degree Celsius overnight Embed the EBs in tissue-Tek OCT compound Cut frozen sections to 4-20 μm thickness

  Primary antibody 1: Rabbit anti cytokeratin 8 (Anti-Cytokeratin 8 antibody [EP1628Y] - Cytoskeleton Marker ab53280), 1:100

  Primary antibody 2: Rat anti-perlecan, 1:100
  Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081), 1:200

  Secondary antibody 2: Goat polyclonal Secondary Antibody to Rat IgG - H&L (Cy5®) pre-adsorbed, 1:200
  Nuclei were counterstained with DAPI

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HL-60 cells labelling S100A8+S100A9 with Anti-S100A8 + S100A9 antibody [RM1038] ab288715 at 1/500 (0.96 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing membranous staining in HL-60 cell line is observed. 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 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

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  Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse PBMC cells labelling S100A8+S100A9 with Anti-S100A8 + S100A9 antibody [RM1038] ab288715 at 1/500 (0.96 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing membranous staining in subsets of mouse PBMCs is observed. 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 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Intracellular flow cytometric analysis of 2% paraformaldehyde fixed, 0.1% saponin permeabilised human peripheral blood mononuclear cell (PBMC) treated with 1μg/ml Lipopolysaccharide (LPS) for 22 hours, then add 3uM Monensin for another 2h (Right). Untreated control (Left).

  Primary antibody: Anti-IL-8 antibody [EPR26511-74] ab289967, at 1/500 dilution.

  Secondary antibody: Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution.

  Scatter image shows specific IL-8 expression in LPS induced monocyte population.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast cell line) cells labeling PUS7 with Anti-PUS7 antibody [EPR25172-31] ab289857 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. The nuclear counterstain is DAPI (Blue). Tubulin is labeled using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) at 1/200 dilution (Red).

  Secondary antibody only control: Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed secondary antibody at 1/1000 dilution.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized HL-60 (Human acute promyelocytic leukemia promyeloblast) cells labelling S100A8+S100A9 with Anti-S100A8 + S100A9 antibody [RM1038] ab288715 at 1/500 dilution (0.1ug)(Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse blood cells cells labelling S100A8+S100A9 with Anti-S100A8 + S100A9 antibody [RM1038] ab288715 at 1/500 dilution (0.1 µg)/ Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control. A Goat Anti-Rabbit IgG (Alexa Fluor 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescence staining of GABA Transporter 1 / GAT 1 using Anti-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins 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-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 at 1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling MAP2 with Anti-MAP2 antibody [RM1037] - Neuronal Marker ab288714 at 1/500 (0.942 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing positive staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 ?m followed by image processing with maximum Z projection. is observed. Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/ml dilution.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Parental HeLa (Human cervix adenocarcinoma epithelial cell, Right) / SIRT6 KO HeLa (Left) cells labelling SIRT6 with Anti-SIRT6 antibody [EPR26255-85] ab289970 at 1/5000 dilution (0.01μg) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Positive staining on HeLa cells (ab255448), while no staining on SIRT6 knockout HeLa cells (Human SIRT6 knockout HeLa cell line ab265054).

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-N-BE(2) cells labelling MAP2 with Anti-MAP2 antibody [RM1037] - Neuronal Marker ab288714 at 1/500 (0.942 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 µg/ml dilution (Green). Confocal image showing cytoplasmic staining in SK-N-BE(2) cell line is observed. 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 2.5 µg/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 µg/ml dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescence staining of GABA Transporter 1 / GAT 1 using Anti-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 mins 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-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 at 5 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

  Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized SIRT1 KO A549(Human SIRT1 knock out human lung carcinoma epithelial cell, Green) / A549(Megenta) cells labelling SIRT1 with Anti-SIRT1 antibody [19A7AB4] ab110304 at 1/1000 dilution (0.1µg)/Red followed by a Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at a 1/5000 dilution (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black and Grey) isotype control.

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of Neuro-2a (mouse neuroblastoma neuroblast, Left) / RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage, Right) cells labelling CD18 with Anti-CD18 antibody [EPR29884-566] ab324101 at 1/5000 dilution (0.01ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

  Low expression: Neuro-2a.
  
  Gated on viable cells.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescence staining of SOX9 using Anti-SOX9 antibody [EPR14335] ab185230 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins 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-SOX9 antibody [EPR14335] ab185230 at 5 μg/ml, Anti-GFAP antibody - Astrocyte Marker ab4674 (anti-GFAP) at 1/1000 dilution and Anti-NeuN antibody [1B7] - Neuronal Marker ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
  As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative.
  Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Histone H3 with Anti-Histone H3 antibody [RM1288] - Nuclear Marker ab322707 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse lung (fresh frozen) tissue labeling Agrin with Anti-Agrin antibody [RM2082] ab324022 at 1/50 (9.98 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

  Confocal image showing positive staining in endothelial cells on mouse lung. The nuclear counterstain was DAPI (Blue). The section was incubated with Anti-Agrin antibody [RM2082] ab324022 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidney epithelial cell) cells labelling OSMR with Anti-OSMR antibody [RM2085] ab324099 at 1/50 (9.94 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing positive staining in 293T cells transfected with a human OSMR expression vector containing a myc-His-tag® (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Myc-Tag Mouse mAb (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse splenocyte cells labelling Neogenin with Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Low expression: confocal image showing no staining in mouse splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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

  -ve control 1: Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
  
  -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Neogenin with Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing positive staining in mouse primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  -ve control 1: Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
  
  -ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-OV-3(human ovarian cancer epithelial cell) cells labelling PINK1 with Anti-PINK1 antibody [EPR29146-340] ab323807 at 1/50 (10.34 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 g/ml dilution (Green).

  Confocal image showing mitochondrial staining in SK-OV-3 cells (shown in green) treatment with 30 μM CCCP for 6 hours. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 g/ml dilution.

  anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Counterstain Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 2 g/ml dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescence staining of SOX9 using Anti-SOX9 antibody [EPR14335-78] ab185966 in primary hippocampal mouse neurons/glia, (obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP), DIV14. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins 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-SOX9 antibody [EPR14335-78] ab185966 at 1 μg/ml, Anti-GFAP antibody - Astrocyte Marker ab4674 (anti-GFAP) at 1/1000 dilution and Anti-NeuN antibody [1B7] - Neuronal Marker ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green), Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) (shown in purple), all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
  As expected, most GFAP positive cells are also SOX9 positive, while NeuN positive cells are SOX9 negative. SOX9 positive cells, which are not GFAP positive (e.g. asterisk) are likely neural stem cells/ oligodendrocyte precursor cells present in the culture.
  Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PINK1 KO HEK-293T (PINK1 knockout human embryonic kidney epithelial cell), Human PINK1 knockout HEK-293T cell line ab266393 cells labelling PINK1 with Anti-PINK1 antibody [EPR29146-340] ab323807 at 1/50 (10.34 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 g/ml dilution (Green).

  Confocal image showing increased mitochondrial staining in wildtype HEK-293T cells treated with 10 μM Valinomycin for 24 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 g/ml dilution.

  anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  Counterstain Secondary antibody only control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 2 g/ml dilution.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized A-172 (human brain glioblastoma cell) cells labelling OSMR with Anti-OSMR antibody [RM2085] ab324099 at 1/50 (9.94 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing membranous and cytoplasmic staining in A-172 and no staining in JAR cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
  
  Negative control: JAR (PMID: 8999038).
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  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 (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

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  Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of KARPAS-299 (human T cell lymphoma cell) (Right) / HepG2 (human hepatocellular carcinoma epithelial cell) (Left) cells labelling HLA DR + DP + DQ with Anti-HLA DR + DP + DQ antibody [CR3/43] ab7856 at 1/1000 dilution (0.1&micor;g)/Red followed by a secondary antibody Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) used at a 1/5000 dilution compared with isotype control Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black and unlabelled control Cell without incubation with primary antibody and secondary antibody / Blue.

  Gated on viable cells.
  Negative control: HepG2.

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  Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling Cofilin with Anti-Cofilin antibody [RM1210] - Loading Control ab319061 at 1/500 dilution (0.1ug) (Magenta) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Frozen sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Positive staining on rat cerebellum.

  Fresh rat cerebellum was fixed with 4% PFA and permeabilised with 0.2 % Triton X100. Anti-pan SCN antibody [EPR25134-4] ab300112 was used as a primary antibody at 1/500 dilution.

  ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was used as a secondary antibody at 1/1000 dilution.

  DAPI was used as a nuclear counterstain.

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  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labelling OSMR with Anti-OSMR antibody [RM2085] ab324099 at 1/50 (9.94 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing mainly membranous staining in A375 and no staining in Jurkat cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
  
  Negative control: Jurkat (PMID: 8999038).
  
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  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 (Magenta). The Nuclear counterstain was DAPI (Blue).

  Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• 

  Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Neogenin with Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

  Confocal image showing positive staining in rat primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

  -ve control 1: Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
  
  -ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.