PRODUCT CODE: EM140707

GFAP Mouse Monoclonal Antibody [1-D4] (EM140707)

  • IVD–IHC

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of GFAP on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM140707, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Rat brain tissue lysate<br />
Lane 2: Human brain tissue lysate
  • Western blot analysis of GFAP on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM140707, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Rat brain tissue lysate<br />
Lane 2: Human brain tissue lysate
  • 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-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • 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 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-GFAP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM140707, 1/400) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue using anti-GFAP antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM140707, 1/400) for 30 minutes 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.
  • 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-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Western blot analysis of GFAP on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM140707, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Rat brain tissue lysate
Lane 2: Human brain tissue lysate

Applications

  • WB

  • IHC-P

  • ICC

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

GFAP Mouse Monoclonal Antibody [1-D4] (EM140707)

Immunogen

Synthetic peptide within c-terminal human gfap.

Host

Mouse

Positive Control

Mouse brain tissue, rat brain tissue, human brain tissue,A172 ,N2A ,Hela.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

1-D4

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

2 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

50 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:2,000-1:5,000

  • ICC:1:200

  • IHC-P

  • 1:200

  • FC

  • 1:200

TARGET

UNIPROT #

PROTEIN NAME

GFAP

SYNONYMS

wu:fb34h11 antibody; ALXDRD antibody; cb345 antibody; etID36982.3 antibody; FLJ42474 antibody; FLJ45472 antibody; GFAP antibody; GFAP_HUMAN antibody; gfapl antibody; Glial fibrillary acidic protein antibody; Intermediate filament protein antibody; wu:fk42c12 antibody; xx:af506734 antibody; zgc:110485 antibody

SEQUENCE SIMILARITIES

Belongs to the intermediate filament family.

TISSUE SPECIFICITY

Expressed in cells lacking fibronectin.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated by PKN1.

SUBCELLULAR LOCATION

Cytoplasm.

FUNCTION

Glial fibrillary acidic protein (GFAP) is a protein that is encoded by the GFAP gene in humans. It is a type III intermediate filament (IF) protein that is expressed by numerous cell types of the central nervous system (CNS), including astrocytes and ependymal cells during development. GFAP has also been found to be expressed in glomeruli and peritubular fibroblasts taken from rat kidneys, Leydig cells of the testis in both hamsters and humans, human keratinocytes, human osteocytes and chondrocytes and stellate cells of the pancreas and liver in rats. GFAP is closely related to the other three non-epithelial type III IF family members, vimentin, desmin and peripherin, which are all involved in the structure and function of the cell’s cytoskeleton. GFAP is thought to help to maintain astrocyte mechanical strength as well as the shape of cells, but its exact function remains poorly understood, despite the number of studies using it as a cell marker. There are multiple disorders associated with improper GFAP regulation, and injury can cause glial cells to react in detrimental ways. Glial scarring is a consequence of several neurodegenerative conditions, as well as injury that severs neural material. Another condition directly related to GFAP is Alexander disease, a rare genetic disorder. Notably, the expression of some GFAP isoforms have been reported to decrease in response to acute infection or neurodegeneration. Additionally, reduction in GFAP expression has also been reported in Wernicke's encephalopathy.