EM140707

GFAP Mouse Monoclonal Antibody [1-D4]

cat.: EM140707

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, IF-Tissue, FC
Clonality:	Monoclonal
Clone number:	1-D4

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 50 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within C-terminal human GFAP.
Positive control:	Human brain tissue lysate, rat brain tissue lysate, human cerebellum tissue lysate, rat cerebellum tissue lysate, A172, N2A, rat brain tissue, rat cerebellum tissue, Hela, rat hippocampus tissue.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:5,000 1:50-1:500 1:50-1:500 1:500-1:1,000 1:50-1:100
Uniprot #:	SwissProt: P14136 Human \| P03995 Mouse \| P47819 Rat
Alternative names:	wu:fb34h11 ALXDRD cb345 etID36982.3 FLJ42474 FLJ45472 GFAP GFAP_HUMAN gfapl Glial fibrillary acidic protein Intermediate filament protein wu:fk42c12 xx:af506734 zgc:110485

Images

	Fig1: Western blot analysis of GFAP on different lysates with Mouse anti-GFAP antibody (EM140707) at 1/5,000 dilution. Lane 1: Human brain tissue lysate Lane 2: Rat brain tissue lysate Lane 3: Human cerebellum tissue lysate Lane 4: Rat cerebellum tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 50 kDa Observed band size: 50 kDa Exposure time: 2 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM140707) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.
	Fig2: ICC staining of GFAP in A172 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM140707, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
	Fig3: ICC staining of GFAP in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (EM140707, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.

	Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Mouse anti-GFAP antibody (EM140707) at 1/600 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM140707) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Mouse anti-GFAP antibody (EM140707) at 1/600 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM140707) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Flow cytometric analysis of GFAP was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (EM140707, 1/100) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
	Fig7: Immunohistochemical analysis of paraffin-embedded human brain tissue with Mouse anti-GFAP antibody (EM140707) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM140707) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig8: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Mouse anti-GFAP antibody (EM140707) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM140707) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig9: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Mouse anti-GFAP antibody (EM140707) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM140707) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig10: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Mouse anti-GFAP antibody (EM140707) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (EM140707) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.