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Purified Mouse Anti-GFAP Cocktail

Purified Mouse Anti-GFAP Cocktail

(RUO)
Purified Mouse Anti-GFAP Cocktail
Immunohistochemical staining of GFAP in human brain cells. A formalin-fixed, paraffin-embedded section of human brain was stained for GFAP using Purified Mouse Anti-GFAP Cocktail (Cat. No. 556330), at a concentration of 10-15 µg/ml.  Samples were counterstained with DAB Substrate Kit (Cat. No. 550880) and hematoxylin, then developed with Anti-Mouse Ig HRP Detection Kit (Cat. No. 551011). The arrow indicates the stained astrocyte.
Purified Mouse Anti-GFAP Cocktail

Flow cytometric analysis of GFAP expression on human brain cell line U373. U373 cells were stained with either Purified Mouse Anti-GFAP Cocktail (solid line histogram) or Purified Mouse IgG2b, κ Isotype Control (dashed line histogram; Cat. No. 556654), followed by PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable cells.

Immunohistochemical staining of GFAP in human brain cells. A formalin-fixed, paraffin-embedded section of human brain was stained for GFAP using Purified Mouse Anti-GFAP Cocktail (Cat. No. 556330), at a concentration of 10-15 µg/ml.  Samples were counterstained with DAB Substrate Kit (Cat. No. 550880) and hematoxylin, then developed with Anti-Mouse Ig HRP Detection Kit (Cat. No. 551011). The arrow indicates the stained astrocyte.

Flow cytometric analysis of GFAP expression on human brain cell line U373. U373 cells were stained with either Purified Mouse Anti-GFAP Cocktail (solid line histogram) or Purified Mouse IgG2b, κ Isotype Control (dashed line histogram; Cat. No. 556654), followed by PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable cells.

Product Details
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BD Pharmingen™
0.5 mg/ml
Human (QC Testing), Mouse, Rat, Pig, Dog, Chicken, Rabbit, Cow, Guinea Pig, Sheep (Tested in Development)
Flow cytometry (Routinely Tested), Immunofluorescence, Immunohistochemistry-paraffin (Tested During Development), Immunohistochemistry-frozen, Western blot (Reported)
RUO
AB_396368
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.


Description

GFAP (Glial Fibrillary Acid Protein) is the major protein of glial filaments in differentiated astrocytes. BD Pharmingen offers a panel of monoclonal antibodies (4A11, 1B4, 2E1) that specifically recognize GFAP. They do not cross-react with other intermediate filaments such as vimentin, neurofilament proteins, desmin, keratin, neurotubules or microfilaments. Bovine spinal cord homogenate was used as immunogen for these clones. Clones 4A11, 1B4, and 2E1 have broad species reactivity, recognizing GFAP in brain homogenates from human, mouse, rat, bovine, ovine, canine, porcine, rabbit, guinea pig and chicken. The cocktail preparation was made by combining all three antibodies in equal concentrations.

Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Applications include indirect immunofluorescence of tissue-cultured cells, immunohistochemical staining of formalin-fixed paraffin-embedded brain tissue sections (10-15 µg/ml); and western blot analysis (1-2 µg/ml). Rat brain is suggested as a positive control. BD Pharmingen also offers these GFAP-specific antibodies separately: clone 4A11 (Cat. No. 556327), clone 1B4 (Cat. No. 556328), clone 2E1 (Cat. No. 556329).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
556330 Rev. 8
Components
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Description Clone Isotype EntrezGene ID
Purified Anti-GFAP 2E1 IgG2b, N/A
Purified Anti-GFAP 1B4 IgG2b, N/A
Purified Anti-GFAP 4A11 IgG2b, N/A
556330 Rev. 8
Citations & References
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Development References (3)

  1. McLendon RE, Bigner DD. Immunohistochemistry of the glial fibrillary acidic protein: basic and applied considerations. Brain Pathol. 1994; 4(3):221-228. (Clone-specific). View Reference
  2. McLendon RE, Burger PC, Pegram CN, Eng LF, Bigner DD. The immunohistochemical application of three anti-GFAP monoclonal antibodies to formalin-fixed, paraffin-embedded, normal and neoplastic brain tissues. J Neuropathol Exp Neurol. 1986; 45(6):692-703. (Clone-specific: Immunohistochemistry). View Reference
  3. Pegram CN, Eng LF, Wikstrand CJ, McComb RD, Lee YL, Bigner DD. Monoclonal antibodies reactive with epitopes restricted to glial fibrillary acidic proteins of several species. Neurochem Pathol. 1985; 3(2):119-138. (Clone-specific: Immunohistochemistry, Western blot). View Reference
556330 Rev. 8

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.