Anti-GFAP Antibody Picoband® (monoclonal, 3F2)

Western blot analysis of GFAP using anti-GFAP antibody (M00213-8).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat brain tissue lysates,
Lane 2: mouse brain tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-GFAP antigen affinity purified monoclonal antibody (Catalog # M00213-8) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for GFAP at approximately 50KD. The expected band size for GFAP is at 50KD.

IHC analysis of GFAP using anti-GFAP antibody (M00213-8).
GFAP was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-GFAP Antibody (M00213-8) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of GFAP using anti-GFAP antibody (M00213-8).
GFAP was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-GFAP Antibody (M00213-8) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

IHC analysis of GFAP using anti-GFAP antibody (M00213-8).
GFAP was detected in a paraffin-embedded section of rat cerebral cortex tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml mouse anti-GFAP Antibody (M00213-8) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

IHC analysis of GFAP using anti-GFAP antibody (M00213-8).
GFAP was detected in a paraffin-embedded section of rat hippocampus tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml mouse anti-GFAP Antibody (M00213-8) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

IHC analysis of GFAP using anti-GFAP antibody (M00213-8).
GFAP was detected in a paraffin-embedded section of mouse hippocampus tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml mouse anti-GFAP Antibody (M00213-8) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.

IF analysis of Histone H3 and GFAP using anti-Histone H3 antibody (A12477-2) and anti-GFAP antibody (M00213-8).
Histone H3 and GFAP was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 μg/mL rabbit anti-Histone H3 antibody (A12477-2) and mouse anti-GFAP antibody (M00213-8) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127), Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

TUDCA promoted neuron regeneration along endogenous NSCs migration at day 7 after SCI. (A) Co-immunofluorescence showed endogenous NSCs (Nestin, green) and reactive astrocytes (GFAP, red) at the margin of the lesion site at day 7 after SCI. (B) Endogenous NSCs (Nestin, green) and neuron (NeuN, red) at the margin of the lesion site at day 7 after SCI. (C, D) Quantitative polymerase chain reaction (qPCR) showing the expression of Nestin, GFAP, NeuN, MAP2 and Oligo 2 at day 7 after SCI. All experiments were performed in triplicated and data were presented means ± SEM, n = 3 per group. *P < 0.05, **P < 0.01.
Index in PubMed under a CC BY license. PMID: 40276612

TUDCA regulated macrophages and reactive astrocytes distribution. Co-immunofluorescence images showed macrophages (CD68, red) and reactive astrocytes (GFAP, green) at day 3 and day 7 after SCI.
Index in PubMed under a CC BY license. PMID: 40276612

TUDCA regulated monocytes distribution and impacted glial scar formation. Co-immunofluorescence images showed reactive astrocytes (GFAP, green) and monocytes (CD11b, red) at day 3 and day 7 after SCI.
Index in PubMed under a CC BY license. PMID: 40276612