Anti-GFAP antibody [EPR1034Y] - BSA and Azide free

30 product images

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  Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Anti-GFAP antibody [EPR1034Y] ab68428 at 1/20 dilution immunoprecipitating GFAP in rat brain whole cell lysate observed at 50 KDa (lanes 1 and 2).
  

  Lane 1 (input): Rat brain whole cell lysate 10ug

  Lane 2 (+): Anti-GFAP antibody [EPR1034Y] ab68428 + Rat brain whole cell lysate

  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-GFAP antibody [EPR1034Y] ab68428 in Rat brain whole cell lysate

  For western blotting, Anti-GFAP antibody [EPR1034Y] ab68428 was used followed by VeriBlot for IP (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) for detection at a dilution of 1/10,000.

  Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

  All lanes: Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] (Anti-GFAP antibody [EPR1034Y] ab68428)

  Predicted band size: 49 kDa

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  Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428, the same antibody clone in a different buffer formulation.
  

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary brain cells cells labelling GFAP with Anti-GFAP antibody [EPR1034Y] ab68428 at 1/500 dilution (0.1ug)/ 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  IHC image of GFAP staining in a formalin fixed, paraffin embedded normal human hippocampus tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with Anti-GFAP antibody [EPR1034Y] ab68428 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

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

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

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  Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural mix culture cells labelling GFAP with Anti-GFAP antibody [EPR1034Y] ab68428 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. ab10062 anti-GFAP antibody [GF5] at 1/100 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2μg/ml) was used as a counterstain. The Nuclear counterstain was DAPI (Blue).
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

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  Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebrum tissue labeling GFAP with Anti-GFAP antibody [EPR1034Y] ab68428 at 1/1000 dilution (2.011 µg/mL) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL) (Green). Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (Blue).

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL).<\p>

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

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  Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebrum tissue labeling GFAP with Anti-GFAP antibody [EPR1034Y] ab68428 at 1/1000 dilution (2.011 µg/mL) followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL) (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

  Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 µg/mL).<\p>

  Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

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  Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-GFAP antibody [EPR1034Y] ab68428).
  

  IHC image of GFAP staining in a section of frozen normal human cerebral cortex performed on a Leica BOND^TM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with Anti-GFAP antibody [EPR1034Y] ab68428, 1/250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

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

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue sections labelling GFAP with purified Anti-GFAP antibody [EPR1034Y] ab68428 at a dilution of 1/500. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP) ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue sections labelling GFAP with purified Anti-GFAP antibody [EPR1034Y] ab68428 at a dilution of 1/500. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP) ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunohistochemical analysis of paraffin-embedded human colon tissue sections labelling GFAP with purified Anti-GFAP antibody [EPR1034Y] ab68428 at a dilution of 1/500. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP) ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. 

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue sections labelling GFAP with purified Anti-GFAP antibody [EPR1034Y] ab68428 at a dilution of 1/500. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP) ab97051, Goat Anti-Rabbit IgG H&L (HRP) at a dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunohistochemical analysis of formalin-fixed paraffin-embedded mouse brain tissue section labelling GFAP with unpurified Anti-GFAP antibody [EPR1034Y] ab68428 at dilution of 1/250.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

  Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunohistochemical analysis of formalin-fixed paraffin-embedded human brain (left) and human glioma (right) tissue sections labelling GFAP with unpurified Anti-GFAP antibody [EPR1034Y] ab68428 at dilution of 1/250.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

  Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of human cerebrum tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] ab68428, gray; Opal™690), anti-Myelin PLP (Anti-Myelin PLP antibody [EPR23504-106] ab254363, green; Opal™520) and anti-MAP2 (Anti-MAP2 antibody [EPR22641-106] ab254263, red; Opal™570) on human cerebrum tissue. Panel B: anti-MAP2 stained cell body and dendrites of neurons. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] ab68428 (1/50 dilution), Anti-Myelin PLP antibody [EPR23504-106] ab254363 (1/2000 dilution), and Anti-MAP2 antibody [EPR22641-106] ab254263 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  
  The data was developed using Anti-GFAP antibody [EPR1034Y] ab68428 , the same antibody clone in a different buffer formulation.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of the Human cerebellum (Formalin/PFA-fixed paraffin-embedded sections).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  Opal Polymer HRP Ms + Rb was used as a secondary antibody.DAPI (blue) was used as a nuclear counter stain.

  Panel A: Merged staining of anti-GFAP (gray; Opal™690), anti-P2Y12 (green; Opal™520) and anti-MAP2 (red; Opal™570) on human cerebellum.
  Panel B: Anti-MAP2 stained cell body and dendrites of neurons.
  Panel C: Anti-P2Y12 stained on microglial cells.
  Panel D: Anti-GFAP stained on astrocytes.

  The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] ab68428, Anti-P2Y12 antibody [EPR23511-72] ab254347, and Anti-MAP2 antibody [EPR22641-106] ab254263 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  The immunostaining was performed on a Leica Biosystems BOND^BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of the Human cerebrum (Formalin/PFA-fixed paraffin-embedded sections).

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  Opal Polymer HRP Ms + Rb was used as a secondary antibody.DAPI (blue) was used as a nuclear counter stain.

  Panel A: Merged staining of anti-GFAP (gray; Opal™690), anti-P2Y12 (green; Opal™520) and anti-MAP2 (red; Opal™570) on human cerebrum.
  Panel B: Anti-MAP2 stained cell body and dendrites of neurons.
  Panel C: Anti-P2Y12 stained on microglial cells.
  Panel D: Anti-GFAP stained on astrocytes.

  The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] ab68428, Anti-P2Y12 antibody [EPR23511-72] ab254347, and Anti-MAP2 antibody [EPR22641-106] ab254263 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse midbrain tissue staining THP2 with Anti-TPH2 antibody [EPR25100-29] ab288067 at a 1/2000 (0.315 ug/ml) dilution, P2Y12 with Anti-P2Y12 antibody [EPR26298-93] ab300140 at 1/40000 (0.013 ug/ml) dilution and GFAP with ab218309 at 1/1000 ( 1.325 ug/ml) dilution.

  Panel A: merged staining of anti-THP2 (green; Opal™520), anti-P2Y12 (grey; Opal™570) and anti-GFAP (magenta; Opal™690) on mouse midbrain.
  Panel B: anti-THP2 staining the serotonergic neurons in mouse midbrain.
  Panel C: anti-P2Y12 staining microglia in mouse midbrain.
  Panel D: anti-GFAP staining astrocytes in mouse midbrain.
  Nuclear DNA was labeled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of ab2888067, Anti-P2Y12 antibody [EPR26298-93] ab300140 and ab218309 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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  Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Immunofluorescence staining of GFAP using Anti-GFAP antibody [EPR1034Y] ab68428 in primary rat hippocampal mixed glia, (prepared from P2 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDPHP4m), DIV4. The cells were fixed with 100% MeOH (5 min), permeabilized with 0.1% Triton-X-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-GFAP antibody [EPR1034Y] ab68428 at 0.1 μg/ml and Anti-GFAP antibody ab4674, Anti-GFAP antibody, at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176, Goat Anti-Chicken IgY 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. The antibody Anti-GFAP antibody [EPR1034Y] ab68428 gave comparable results using 4% formaldehyde fixation (10 min).
  
  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-GFAP antibody [EPR1034Y] ab68428).

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded rat spinal cord tissue sections.

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-GFAP (red; Opal™570) on rat spinal cord.

  Panel B: anti-GPR17 staining oligodendrocytes in rat spinal cord.

  Panel C: anti-P2RY12 staining microglia in rat spinal cord.

  Panel D: anti-GFAP staining astrocytes in rat spinal cord.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at a 1/500 dilution, Anti-P2Y12 antibody [EPR26298-93] ab300140 at a 1/40000 dilution, and ab218309 at a 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  Nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse cerebrum tissue sections.
  

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-GFAP (red; Opal™570) on mouse cerebrum.

  Panel B: anti-GPR17 staining oligodendrocytes in mouse cerebrum.

  Panel C: anti-P2RY12 staining microglia in mouse cerebrum.

  Panel D: anti-GFAP staining astrocytes in mouse cerebrum.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at a 1/500 dilution, Anti-P2Y12 antibody [EPR26298-93] ab300140 at a 1/140000 dilution, and ab218309 at a 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded rat cerebrum tissue sections.
  

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-GFAP (red; Opal™570) on rat cerebrum.

  Panel B: anti-GPR17 staining oligodendrocytes in rat cerebrum.

  Panel C: anti-P2RY12 staining microglia in rat cerebrum.

  Panel D: anti-GFAP staining astrocytes in rat cerebrum.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at 1/500 dilution, Anti-P2Y12 antibody [EPR26298-93] ab300140 at 1/140000 dilution , and ab218309 at 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The secondary was Opal Polymer HRP Ms + Rb and nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428 the same antibody clone in a different buffer formulation.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse cerebellum tissue sections.

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-GFAP (red; Opal™570) on mouse cerebellum.

  Panel B: anti-GPR17 staining oligodendrocytes in mouse cerebellum.

  Panel C: anti-P2RY12 staining microglia in mouse cerebellum.

  Panel D: anti-GFAP staining astrocytes in mouse cerebellum.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at a 1/500 dilution, Anti-P2Y12 antibody [EPR26298-93] ab300140 at a 1/40000 dilution, and ab218309 at a 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  Nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse spinal cord tissue sections.

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-GFAP (red; Opal™570) on mouse spinal cord.

  Panel B: anti-GPR17 staining oligodendrocytes in mouse spinal cord.

  Panel C: anti-P2RY12 staining microglia in mouse spinal cord.

  Panel D: anti-GFAP staining astrocytes in mouse spinal cord.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at a 1/500 dilution, Anti-P2Y12 antibody [EPR26298-93] ab300140 at a 1/40000 dilution, and ab218309 at a 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  Nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded mouse hippocampus tissue sections.

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-P2RY12 (green; Opal™520) and anti-GFAP (red; Opal™570) on mouse hippocampus.

  Panel B: anti-GPR17 staining oligodendrocytes in mouse hippocampus.

  Panel C: anti-P2RY12 staining microglia in mouse hippocampus.

  Panel D: anti-GFAP staining astrocytes in mouse hippocampus.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at a 1/500 dilution, Anti-P2Y12 antibody [EPR26298-93] ab300140 at a 1/40000, and ab218309 at a 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  Nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human cerebrum tissue sections.
  

  Panel A: merged staining of anti-GPR17 (magenta; Opal™690), anti-TMEM119 (green; Opal™520) and anti-GFAP (red; Opal™570) on human cerebrum.

  Panel B: anti-GPR17 staining oligodendrocytes in human cerebrum.

  Panel C: anti-TMEM119 staining microglia in human cerebrum.

  Panel D: anti-GFAP staining astrocytes in human cerebrum.

  The section was incubated in three rounds of staining: in the order of Anti-GPCR GPR17 antibody [EPR26423-34] ab316105 at a 1/500 dilution, Anti-TMEM119 antibody [EPR25865-89] ab306583 at a 1/2000 dilution, and ab218309 at a 1/1000 dilution for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Nuclear counterstaining with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human astrocytoma tissue staining TREM2 with Anti-TREM2 antibody [EPR26209-22] ab318262 at a 1:100 (5.29 ug/ml) dilution, Anti-TMEM119 antibody [EPR25865-89] ab306583 anti-TMEM119 used at 1:2000 (0.255 ug/ml) dilution and ab218309 anti-GFAP used at a 1:1000 (1.325 ug/ml) dilution.

  Panel A: merged staining of anti-TREM2 (green; Opal™520), anti-TMEM119 (magenta; Opal™690) and anti-GFAP (yellow; Opal™570) on human astrocytoma.
  
  Panel B: anti-TREM2 staining microglia in human astrocytoma.
  
  Panel C: anti-TMEM119 staining microglia in human astrocytoma.
  
  Panel D: anti-GFAP staining astrocyte in human astrocytoma.
  
  Nuclear DNA was labeled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-TREM2 antibody [EPR26209-22] ab318262, Anti-TMEM119 antibody [EPR25865-89] ab306583 and ab218309 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  Nuclear counter stain with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human cerebrum tissue staining TREM2 with Anti-TREM2 antibody [EPR26209-22] ab318262 at a 1:100 (5.29 ug/ml) dilution, Anti-TMEM119 antibody [EPR25865-89] ab306583 anti-TMEM119 used at 1:2000 (0.255 ug/ml) dilution and ab218309 anti-GFAP used at a 1:1000 (1.325 ug/ml) dilution.

  Panel A: merged staining of anti-TREM2 (green; Opal™520), anti-TMEM119 (magenta; Opal™690) and anti-GFAP (yellow; Opal™570) on human cerebrum.
  
  Panel B: anti-TREM2 staining microglia in human cerebrum.
  
  Panel C: anti-TMEM119 staining microglia in human cerebrum.
  
  Panel D: anti-GFAP staining astrocytes in human cerebrum.
  
  Nuclear DNA was labeled with DAPI (shown in blue).

  The section was incubated in three rounds of staining: in the order of Anti-TREM2 antibody [EPR26209-22] ab318262, Anti-TMEM119 antibody [EPR25865-89] ab306583 and ab218309 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

  The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

  Nuclear counter stain with DAPI.

  Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of the human cerebellum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] ab68428, gray; Opal™690), anti-Myelin PLP (Anti-Myelin PLP antibody [EPR23504-106] ab254363, green; Opal™520) and anti-P2Y12 (Anti-P2Y12 antibody [EPR23511-72] ab254347 red; Opal™570) on human cerebellum. Panel B: anti-P2Y12 stained on microglial cells. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] ab68428 (1/50 dilution), Anti-Myelin PLP antibody [EPR23504-106] ab254363 (1/2000 dilution), and Anti-P2Y12 antibody [EPR23511-72] ab254347 (1/1000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  
  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428, the same antibody clone in a different buffer formulation.

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  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of the human cerebrum (Formalin/PFA-fixed paraffin-embedded sections). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] ab68428, gray; Opal™690), anti-Myelin PLP (Anti-Myelin PLP antibody [EPR23504-106] ab254363, green; Opal™520) and anti-P2Y12 (Anti-P2Y12 antibody [EPR23511-72] ab254347, red; Opal™570) on human cerebrum. Panel B: anti-P2Y12 stained on microglial cells. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] ab68428 (1/50 dilution), Anti-Myelin PLP antibody [EPR23504-106] ab254363 (1/2000 dilution), and Anti-P2Y12 antibody [EPR23511-72] ab254347 (1/1000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  
  
  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428, the same antibody clone in a different buffer formulation.

• 

  Multiplex immunohistochemistry - Anti-GFAP antibody [EPR1034Y] - BSA and Azide free (ab218309)

  Fluorescence multiplex immunohistochemical analysis of human cerebellum tissue (formalin/PFA-fixed paraffin-embedded section). Panel A: merged staining of anti-GFAP (Anti-GFAP antibody [EPR1034Y] ab68428, gray; Opal™690), anti-Myelin PLP (Anti-Myelin PLP antibody [EPR23504-106] ab254363, green; Opal™520) and anti-MAP2 (Anti-MAP2 antibody [EPR22641-106] ab254263, red; Opal™570) on human cerebellum tissue. Panel B: anti-MAP2 stained cell body and dendrites of neurons. Panel C: anti-Myelin PLP stained on myelin sheaths of oligodendrocytes. Panel D: anti-GFAP stained on astrocytes. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND^®RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining: in the order of Anti-GFAP antibody [EPR1034Y] ab68428 (1/50 dilution), Anti-Myelin PLP antibody [EPR23504-106] ab254363 (1/2000 dilution), and Anti-MAP2 antibody [EPR22641-106] ab254263 (1/4000 dilution) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope.

  
  This data was developed using Anti-GFAP antibody [EPR1034Y] ab68428, the same antibody clone in a different buffer formulation.