Revision 5

#95717Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

50

Source/Isotype:

Mouse IgG2b

UniProt ID:

#P14136

Entrez-Gene Id:

2670

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:100 - 1:200
Immunofluorescence (Immunocytochemistry) 1:100 - 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

GFAP (E8S7G) Mouse mAb recognizes endogenous levels of total GFAP protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with native GFAP purified from pig spinal cord.

Background

The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Major types of intermediate filaments are specifically expressed in particular cell types: cytokeratins in epithelial cells, glial fibrillary acidic protein (GFAP) in glial cells, desmin in skeletal, visceral, and certain vascular smooth muscle cells, vimentin in cells of mesenchymal origin, and neurofilaments in neurons. GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape (1). In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes (2). In addition, GFAP intermediate filaments are also present in nonmyelin-forming Schwann cells in the peripheral nervous system (3).

  1. Eng, L.F. et al. (2000) Neurochem. Res. 25, 1439-51.
  2. Goebel, H.H. et al. (1987) Acta. Histochem. Suppl. 34, 81-93.
  3. Jessen, K.R. et al. (1990) Development 109, 91-103.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#95717

GFAP (E8S7G) Mouse mAb

Western Blotting Image 1: GFAP (E8S7G) Mouse mAb Expand Image
Western blot analysis of extracts from various tissues and cell lines using GFAP (E8S7G) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: GFAP (E8S7G) Mouse mAb Expand Image
Immunoprecipitation of GFAP protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Mouse (E7Q5L) mAb IgG2b Isotype Control #53484, and lane 3 is GFAP (E8S7G) Mouse mAb. Western blot analysis was performed using GFAP (E4L7M) XP® Rabbit mAb #80788. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunoprecipitation Image 2: GFAP (E8S7G) Mouse mAb Expand Image
Immunoprecipitation of GFAP protein from mouse brain tissue extracts. Lane 1 is 10% input, lane 2 is Mouse (E7Q5L) mAb IgG2b Isotype Control #53484, and lane 3 is GFAP (E8S7G) Mouse mAb. Western blot analysis was performed using GFAP (E8S7G) Mouse mAb. Rabbit Anti-Mouse IgG (Light Chain Specific) (D3V2A) mAb (HRP Conjugate) #58802 was used as the secondary antibody.
Immunofluorescence Image 1: GFAP (E8S7G) Mouse mAb Expand Image
Confocal immunofluorescent analysis of rat cerebellum using GFAP (E8S7G) Mouse mAb (red) and UCHL1 (D3T2E) XP® Rabbit mAb #13179 (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #78060 (cyan pseudocolor) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: GFAP (E8S7G) Mouse mAb Expand Image
Confocal immunofluorescent analysis of rat hippocampus using GFAP (E8S7G) Mouse mAb (red) and UCHL1 (D3T2E) XP® Rabbit mAb #13179 (green). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using Iba1/AIF-1 (E4O4W) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #78060 (cyan pseudocolor) and ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: GFAP (E8S7G) Mouse mAb Expand Image
Confocal immunofluorescent analysis of U-251 MG cells (left, positive) or HeLa cells (right, negative) using GFAP (E8S7G) Mouse mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).