S-100 Protein (alpha chain) antibody

Antigen: S-100 Protein (alpha chain)

Clonality: Polyclonal

Host: Rabbit

Reactivity: Mouse (Murine), Human, Rat (Rattus)

Application: Immunohistochemistry (IHC), ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN129527

Additional Information

Alternative name: S-100 Protein

Gene ID: BCBOIA (Bovine), AAP35584 (Human)

Immunogen: This Protein A purified antibody was prepared from whole rabbit serum produced by repeated immunizations with full-length bovine S100 protein (mixture of alphaalpha homodimers and alphabeta heterodimers).

Format: Liquid (sterile filtered)

Description: S-100 protein derived from brain tissue is an acidiccalcium-binding protein with molecular weight of about 21kDa. Inhuman brain tissue S-100 protein is mainly presented as twoisoforms - betabeta homodimer (S-100b) and alphabeta heterodimer (S-100a). Because of its predominant location in astroglial cells S-100 proteincan be used as a sensitive and reliable marker for central nervoussystem injury. Structural damage of glial cells causes leakage of S-100 protein into the extracellular matrix and into cerebrospinal fluid,further releasing into the bloodstream. Measurements of S-100protein in patient serum samples are useful in monitoring oftraumatic brain injury, ischemic brain damage after circulatoryarrests, and in diagnosis and prognosis of clinical outcome in acutestroke. Although predominant among the water-soluble brainproteins, S-100 is also found in a variety of other tissues. S-100 isan intracellular protein that weakly binds calcium. It binds zinc verytightly, however, and this appears to increase the affinity of theprotein for calcium. Distinct binding sites, with different affinities,exist for both ions on each monomer. Physiological concentrationsof potassium ion antagonize the binding of both divalent cations,especially affecting high-affinity calcium-binding sites.

Specificity: This Protein A purified antibody is directed against bovine S100 protein. The product was purified frommonospecific antiserum by Protein A chromatography. A BLAST analysis was used to suggest reactivity with this protein from bovinebased on 100% homology with the immunogen sequence. A 98% homology is noted for S100 alpha chain from primate sources.Mouse, rat and dog show 94% homology with the bovine S100 alpha sequence. Expect cross reactivity with S100 from mostmammalian sources. Cross reactivity with S100 from other specific sources has not been determined. Homologies for the S100 betachain are similar.

Application Details

Application Notes: This Protein A purified antibody has been tested for use in ELISA and western blot. Specific conditions forreactivity should be optimized by the end user. Expect a band ~ 11 kDa in size corresponding to S100 monomer by western blotting inthe appropriate cell lysate or extract. Dilutions: ELISA 1:5,000 - 1:25,000 WESTERN BLOT 1:500 - 1:3,000 IMMUNOHISTOCHEMISTRY 1:200 - 1:2,000 OTHER APPLICATIONS User Optimized

Concentration: 5.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at 4° C prior to opening. Restore with 0.1 ml of deionized water (or equivalent). Dilute only prior toimmediate use. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Expirationdate is one (1) year from date of restoration.

Research Area: Cell Cycle, Cancer

Restrictions: For Research Use only

Images

Western blot using antibodies-online's Affinity Purified anti-S-100 antibody shows detection of a band ~11 kDa corresponding to bovine S-100 monomer (100 ng loaded, arrowhead lane 1). The antibody also detects S-100 from rat brain lysate (lane 2). Approximately 35 ug of a rat brain whole cell lysate was separated by 16% SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:1,000 for 2h at room temperature followed by washes and reaction with a 1:10,000 dilution of IRDye

Immunohistochemistry. Rabbit anti-S-100 protein was used at a 1:500 dilution to detect S-100 by immunohistochemistry using a 2-step indirect method. Dark nuclear staining is observed within basket cells located near the Purkinje cells in the cerebellum. Mouse brain tissue was immersed for 24 hours in 10% neutral buffered formalin and paraffin processed followed by sectioning at 4 microns. No antigen unmasking (HIER) or protease digestion was performed prior to immunostaining. Sections were deparaffinized in xylene, and hydrated through graded alcohol to distilled water. All incubations were done at room temperature. All rinses were either distilled water or Tris-HCl with 0.05% Tween 20. Endogenous peroxidase activity was blocked with 3% Hydrogen peroxide for 10'. Non-specific binding was blocked using PowerBlock (Biogenex) for 10'. Primary antibody was diluted as stated and reacted for 30' followed by washes and the addition of donkey anti-rabbit HRP diluted 1:500 for 30'. DAB+ (Dakocytomation) was used as a substrate and was allowed to react for 5'. Personal Communication, Teri Johnson, Stowers Institute, Kansas City, MO.