S100 Protein Antibody

SKU: orb344628

Description

S100 Protein antibody

Images & Validation

• Tested Applications: ELISA, IHC, WB
• Dilution range: ELISA: 1:5,000 - 1:25,000, IHC: 1:200 - 1:2,000, WB: 1:500 - 1:3,000
• Reactivity: Bovine, Human, Mouse, Rat
• Application Notes:
  This Protein A purified antibody has been tested for use in immunohistochemistry, and western blot and is suitable for ELISA. Specific conditions for reactivity should be optimized by the end user. Expect a band ~ 11 kDa in size corresponding to S100 monomer by western blotting in the appropriate cell lysate or extract.

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: This Protein A purified antibody was prepared from whole rabbit serum produced by repeated immunizations with full-length bovine S100 protein (mixture of aa homodimers and ab heterodimers).
• Purity: This Protein A purified antibody is directed against bovine S100 protein. The product was purified from monospecific antiserum by Protein A chromatography. A BLAST analysis was used to suggest reactivity with this protein from bovine based 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 most mammalian sources. Cross reactivity with S100 from other specific sources has not been determined. Homologies for the S100 beta chain are similar.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Store vial at 4° C prior to restoration. For extended storage aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Concentration: 5.0 mg/mL
• Disclaimer: For research use only

Alternative Names

rabbit anti-S100 Protein Antibody, S100 calcium-binding protein A1, S-100 protein subunit alpha, S-100 protein alpha chain

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'.

Rabbit anti-S100 was used at a 1:500 dilution to detect S100 by immunohistochemistry in human brain astrocyte tumor tissue. Tissue was formalin-fixed and paraffin embedded.

Western blot using Biorbyt'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 µg of a rat brain whole cell lysate (p/n orb348727) was separated by 16% SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:1000 for 2h at room temperature followed by washes and reaction with a 1:10000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature.

Quick Database Links

UniProt

P02639

RefSeq (Protein):NP_001092512.1