Anti-GFAP antibody [EP672Y] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

Immunofluorescence staining of GFAP using ab33922 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 ab33922 at 0.2 μg/ml and ab4674, Anti-GFAP antibody, 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 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 ab33922 gave comparable results using 4% formaldehyde fixation (10 min). This data was developed using ab33922, the same antibody clone in a different buffer formulation.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

This data was developed using ab33922, the same antibody clone in a different buffer formulation.

Immunocytochemistry analysis of Embryonic mouse primary neural cells labeling GFAP with purified ab33922 at 1 : 100 dilution (10 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor^© 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. Cells were counterstained with Tau Mouse mAb 1 : 100 (5 μg/ml). ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) was used as the secondary antibody for the counter stain at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain.

Confocal image showing positive staining in glial cells.

• Mass Cytometry

Collaborator

Mass Cytometry - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

Imaging Mass Cytometry™ (IMC™) image of human glioblastoma brain cancer tissue stained with Anti-GFAP antibody [EP672Y]. ab220820 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm's protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.

Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada

This image is courtesy of the Single Cell & Imaging Mass Cytometry Analysis Platform, Goodman Cancer Research Centre, McGill University

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

This data was developed using ab33922, 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 ab33922 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

This data was developed using ab33922, the same antibody clone in a different buffer formulation.

ICC image of ab33922 stained Rat primary mixed astrocytes culture. The cells were 100% Paraformaldehyde fixed and then incubated in 10% Serum / 0.1M PBS with 10% Donkey serum for 4h. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution.

This image is courtesy of an Abreview submitted by Ruma Raha-Chowdhury.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

This data was developed using ab33922, the same antibody clone in a different buffer formulation.

ab33922 showing positive staining in human Astrocytoma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

This data was developed using ab33922, the same antibody clone in a different buffer formulation.

ab33922 showing negative staining in human Meningioma tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

This data was developed using ab33922, the same antibody clone in a different buffer formulation.

ab33922 showing positive staining in Normal human brain tissue.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-GFAP antibody [EP672Y] - BSA and Azide free (AB220820)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.