Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric)

7 product images

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  Immunocytochemistry - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290)

  Immunocytochemical analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized mouse neural/glia tissue labeling GFAP with ab279290 at 1/50 (16 μg/ml) dilution followed by ab98736 Goat Anti-Mouse IgG (DyLight® 488) pre-adsorbed at 1/1000 dilution (Green). Positive staining on mouse primary astrocytes is observed. The nuclear counterstain was DAPI (Blue).
  

  Counterstain: Anti-GFAP antibody [EPR19996] ab207165 anti-GFAP rabbit mAb at 1/1000 dilution with secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution.

  Negative controls: ab279290 with secondary Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at 1/500 dilution; and Anti-GFAP antibody [EPR19996] ab207165 at 1/1000 dilution with secondary ab98376 at 1/1000 dilution

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  Western blot - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  The molecular weight observed is consistent with the literature (PMID: 824020, 2294, 6340792).

  Exposure times: Lane 1: 3.25 seconds; Lane 2: 48 seconds; Lane 3: 10 seconds.

  All lanes: Western blot - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290) at 1/1000 dilution

  Lane 1: Human brain tissue lysate at 20 µg

  Lane 2: Mouse brain tissue lysate at 20 µg

  Lane 3: Rat brain tissue lysate at 20 µg

  Secondary

  All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

  Predicted band size: 49 kDa

  Observed band size: 40-50 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290)

  IHC image of GFAP staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex performed on a Leica BOND^TM 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 20mins. The section was then incubated with ab279290, 1μg/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG2a, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit 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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  Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290)

  GFAP was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 μg with ab279290 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279290 at 1/1000 dilution. mouse IgG for IP (HRP) (Anti-mouse IgG for IP (HRP) ab131368) was used at 1/5000 dilution.
  

  Lane 1: Rat brain tissue lysate 10μg.

  Lane 2: ab279290 IP in rat brain tissue lysate.

  Lane 3: Mouse monoclonal IgG2a (Mouse IgG2a, Kappa Monoclonal [MOPC-173] -Isotype Control - ChIP Grade ab18413) instead of ab279290 in rat brain tissue lysate.

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

  Exposure time: 23 seconds.

  All lanes: Immunoprecipitation - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290)

  Predicted band size: 49 kDa

  Observed band size: 50 kDa

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  Flow Cytometry (Intracellular) - Anti-GFAP antibody [EPR1034Y] – Mouse IgG2a (Chimeric) (ab279290)

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized rat primary neural/glia cells labelling GFAP with ab279290 at 1/1000 dilution (0.1μg)/ Right compared with a Mouse monoclonal IgG isotype control/ Left. Goat Anti-Mouse IgG (Alexa Fluor^® 647, Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119) at 1/2000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen rat cerebrum tissue labeling GFAP with ab279290 at 1/200 (10 µg/ml) dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 (2μg/mL) dilution. Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (blue).

  Secondary antibody control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

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  Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290)

  Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen mouse cerebrum tissue labeling GFAP with ab279290 at 1/200 (10 µg/ml) dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 (2μg/mL) dilution. Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (blue).

  Secondary antibody control: Secondary antibody is Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.