Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker

21 product images

• 

  Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Western blot staining using rabbit Anti-160 kD Neurofilament Medium antibody

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

  The molecular weight is consistent to what has been described in the literature (PMID:19239416, PMID:27000625).

  Negative control: liver (PMID:30541916).

  Exposure time: 10 seconds.

  All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348) at 1/1000 dilution

  Lane 1: Human cerebellum tissue lysate at 20 µg

  Lane 2: Human nerve tissue lysate at 20 µg

  Lane 3: Human liver tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Predicted band size: 102 kDa

  Observed band size: 160 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labelling 160 kD Neurofilament Medium with ab254348 at 1/4000 (0.111 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab254348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
  

  Positive staining on human cerebellum.

  Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Immunohistochemical analysis of paraffin-embedded mouse liver tissue labelling 160 kD Neurofilament Medium with ab254348 at 1/4000 (0.111 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab254348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
  

  Negative control: No staining on mouse liver.

  Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Immunofluorescent analysis of parental HEK-293T (EDWT04) and NEFM KO HEK-293T (ED040746) cells labelling 160 kD Neurofilament Medium with ab254348 at 1/50 (8.86 μg/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). 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 (Blue).
  

  Confocal image showing cytoplasmic staining in parental HEK-293T cells and no staining in NEFM KO HEK-293T cells.

  Secondary antibody only 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.

• 

  Immunoprecipitation - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium was immunoprecipitated from 0.35 mg mouse brain tissue lysate 10 μg with ab254348 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab254348 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Mouse brain tissue lysate 10 μg.

  Lane 2: ab254348 IP in mouse brain tissue lysate.

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab254348 in mouse brain tissue lysate.

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

  Exposure time: 3.25 secs.

  All lanes: Immunoprecipitation - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  Predicted band size: 102 kDa

  Observed band size: 160 kDa

• 

  Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunofluorescent analysis of mouse primary neural/glia cells fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100, labelling 160 kD Neurofilament Medium with ab254348 at 1/50 (8.86 μg/ml) dilution. Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4 μg/ml). Followed by secondary Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 μg/ml) (Green). Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) was used as secondary counterstain at 1/1000 (2 μg/ml) (Red). The nuclear counterstain was DAPI (Blue).
  

  Confocal image showing cytoplasmic staining in mouse primary neuron cells.
  Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

  Negative Control 1: Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 1/1000 (2 μg/ml)

  Negative Control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 1/500 (4 μg/ml), secondary: Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 1/1000 (2μg/ml)

• 

  Immunocytochemistry/ Immunofluorescence - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunofluorescent analysis of rat primary neural/glia cells fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100, labelling 160 kD Neurofilament Medium with ab254348 at 1/50 (8.86 μg/ml) dilution. Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain at 1/500 (4 μg/ml). Followed by secondary Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 (2 μg/ml) (Green). Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) was used as secondary counterstain at 1/1000 (2 μg/ml) (Red). The nuclear counterstain was DAPI (Blue).
  

  Confocal image showing cytoplasmic staining in rat primary neuron cells.
  Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.

  Negative Control 1: Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 1/1000 (2 μg/ml)

  Negative Control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 1/500 (4 μg/ml), secondary: Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 1/1000 (2 μg/ml)

• 

  Flow Cytometry (Intracellular) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Flow Cytometry (Intracellular) staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized wild-type HEK-293T (human embryonic kidney epithelial cell, Right)/ 160 kD Neurofilament Medium knockout HEK-293T cells (Left) labelling 160 kD Neurofilament Medium with ab254348 at 1/500 dilution (0.1 μg). 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.
  

  Positive staining on human wild-type HEK-293T cell line (ab255449), while no staining on human NEFM knockout HEK-293T cells (Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line ab266741).

• 

  Immunohistochemistry (Frozen sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling 160 kD Neurofilament Medium with ab254348 at 1/100 (4.43 μg/ml) dilution 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 (Green). The nuclear counterstain was DAPI (Blue).
  

  Positive staining on mouse cerebrum is observed.

  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.

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

• 

  Immunohistochemistry (Frozen sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver tissue labeling 160 kD Neurofilament Medium with ab254348 at 1/100 (4.43 μg/ml) dilution 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 (Green). The nuclear counterstain was DAPI (blue).
  

  Negative control: No staining on mouse liver (PMID: 30541916) is observed.

  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.

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

• 

  ELISA - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  ELISA analysis using ab254348 at a range of 0-1000 ng/ml followed by a Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L）at 1/2500 dilution. Antigen concentration: 1000 ng/ml.
  

  Antigens: Mouse 160 kD Neurofilament Medium.

• 

  Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Western blot staining using rabbit Anti-160 kD Neurofilament Medium antibody

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

  The molecular weight is consistent to what has been described in the literature (PMID:19239416, PMID:27000625).

  Negative control: liver (PMID:30541916).

  Exposure times: Lane 1-3: 3.25 secs; Lane 4-6: 10 secs.

  All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Mouse cerebellum tissue lysate at 20 µg

  Lane 3: Mouse liver tissue lysate at 20 µg

  Lane 4: Rat brain tissue lysate at 20 µg

  Lane 5: Rat cerebellum tissue lysate at 20 µg

  Lane 6: Rat liver tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 102 kDa

  Observed band size: 160 kDa

• 

  Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Western blot staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Blocking and diluting buffer and concentration: Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.
  

  Lanes 1-3: Merged signal (red and green). Green - ab254348 observed at 160 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

  ab254348 Anti-NEFM antibody [EPR23510-76] was shown to specifically react with NEFM in wild-type HEK-293T cells. Loss of signal was observed when the knockout cell line Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell line ab266741 (knockout cell lysate Human NEFM (160 kD Neurofilament Medium) knockout HEK-293T cell lysate ab257103) was used. Wild-type and NEFM knockout samples were subjected to SDS-PAGE. ab254348 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348) at 1/1000 dilution

  Lane 1: Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

  Lane 2: NEFM knockout HEK-293T whole cell lysate at 20 µg

  Lane 3: A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution

  Predicted band size: 102 kDa

  Observed band size: 160 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labelling 160 kD Neurofilament Medium with ab254348 at 1/4000 (0.111 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab254348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
  

  Positive staining on human cerebrum.

  Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labelling 160 kD Neurofilament Medium with ab254348 at 1/4000 (0.111 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab254348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
  

  Positive staining on mouse cerebellum.

  Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunoprecipitation - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 μg with ab254348 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab254348 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
  

  Lane 1: Rat brain tissue lysate 10 μg.

  Lane 2: ab254348 IP in rat brain tissue lysate.

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab254348 in rat brain tissue lysate.

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

  Exposure time: 5.5 secs.

  All lanes: Immunoprecipitation - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  Predicted band size: 102 kDa

  Observed band size: 160 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labelling 160 kD Neurofilament Medium with ab254348 at 1/4000 (0.111 μg/ml) dilution followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). The section was incubated with ab254348 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.
  

  Positive staining on rat cerebellum.

  Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• 

  Immunohistochemistry (Frozen sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum tissue labeling 160 kD Neurofilament Medium with ab254348 at 1/100 (4.43 μg/ml) dilution 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 (Green). The nuclear counterstain was DAPI (Blue).
  

  Positive staining on rat cerebrum is observed.

  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.

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

• 

  Immunohistochemistry (Frozen sections) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (ab254348)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver tissue labeling 160 kD Neurofilament Medium with ab254348 at 1/100 (4.43 μg/ml) dilution 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 (Green). The nuclear counterstain was DAPI (blue).
  

  Negative control: No staining on rat liver (PMID: 30541916) is observed.

  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.

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

• 

  Flow Cytometry (Intracellular) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Flow Cytometry (Intracellular) staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized mouse primary neuron cells labelling 160 kD Neurofilament Medium with ab254348 at 1/500 dilution (0.1 μg)/ Right compared with a Rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left.
  

  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.

• 

  Flow Cytometry (Intracellular) - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] - Neuronal Marker (ab254348)

  160 kD Neurofilament Medium Flow Cytometry (Intracellular) staining using rabbit Anti-160 kD Neurofilament Medium antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized rat primary neuron cells cells labelling 160 kD Neurofilament Medium with ab254348 at 1/500 dilution (0.1 μg)/ Right compared with a Rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left.
  

  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.