Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

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Anti-GFAP antibody [EPR19996] (ab207165)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19996] to GFAP
Suitable for: ICC, IHC-P, WB, IHC-Fr, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-GFAP antibody [EPR19996]
See all GFAP primary antibodies
Description

Rabbit monoclonal [EPR19996] to GFAP
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC	Mouse
IHC-Fr	Mouse
IHC-P	Mouse Rat Human
IP	Human
WB	Mouse Rat

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human, mouse and rat brain lysates; rat cerebellum lysate. IHC-P: Human cerebrum and glioma tissues; mouse cerebrum and colon tissues; rat hippocampus and colon tissues. IHC-Fr: Mouse cerebrum tissue. IP: Human brain lysate. ICC: Mouse primary neural/glia cells
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19996
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on glial cells of human cerebrum [PMID: 15378652] is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry - Anti-GFAP antibody [EPR19996] (ab207165)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labelling GFAP with ab207165 at 1/1000 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (2 µg/mL) (Green). Confocal image showing cytoplasmic staining in mouse primary astrocyte. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (4 µg/mL) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 µg/mL) (Red). The Nuclear counterstain was DAPI (Blue).
Immunohistochemistry (Frozen sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling GFAP with ab207165 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Cytoplasmic and membrane staining on glial cells of mouse cerebrum is observed.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Western blot - Anti-GFAP antibody [EPR19996] (ab207165)

Lane 1 : Anti-GFAP antibody [EPR19996] (ab207165) at 1/10000 dilution
Lane 2 : Anti-GFAP antibody [EPR19996] (ab207165) at 1/1000 dilution

Lane 1 : Human brain lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 49 kDa
Observed band size: 40-54 kDa
why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with the literature.

Negative control: NIH/3T3 (PMID: 824020; PMID:2294; PMID: 6340792; PMID: 9466565).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on tumor cells of human glioma is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on glial cells of mouse cerebrum is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Specific cytoplasmic staining on myenteric nerve plexus, and negative on epithelial cells and smooth muscle cells of mouse colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and membrane staining on glial cells of rat hippocampus is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFAP antibody [EPR19996] (ab207165)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling GFAP with ab207165 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Specific cytoplasmic staining on myenteric nerve plexus, and negative on epithelial cells and smooth muscle cells of rat colon is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-GFAP antibody [EPR19996] (ab207165)

All lanes : Anti-GFAP antibody [EPR19996] (ab207165) at 1/5000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : Rat cerebellum lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 49 kDa
Observed band size: 40-54 kDa why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with the literature (PMID: 824020; PMID:2294; PMID: 6340792).
Immunoprecipitation - Anti-GFAP antibody [EPR19996] (ab207165)

GFAP was immunoprecipitated from 0.35 mg of human brain lysate with ab207165 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab207165 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Human brain lysate 10µg (Input).

Lane 2: ab207165 IP in human brain lysate.

Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207165 in human brain lysate.

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

Exposure time: 1 second.
Anti-GFAP antibody [EPR19996] (ab207165)

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