Nestin Antibody

ELISA results of purified Rabbit anti-Nestin Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1 µg of conjugate. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gel, Goat anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n orb347654) and TMB ELISA Peroxidase Substrate (p/n orb348651).

Immunohistochemistry with Anti-nestin antibody at 40X (left) and 20X (right) Tissue: Brain and Cerebellum (right) Fixation: FFPE buffered formalin 10% conc Antigen retrieval: Heat, Citrate pH6.2. Pressure Cooker Primary antibody: 20 ug/ml 1 hour @ room T Secondary antibody: Goat anti Rabbit Polymer HRP Prediluted by the manufacturer 30 min. @ room T Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Immunohistochemistry with anti-nestin antibody showing nestin staining in cytoplasm of ductal epithelium of human salivary gland (B) and in nucleus and cytoplasm of human breast tissue (C). Formalin fixed/paraffin embedded sections were subjected to heat induced epitope retrieval (HIER) at pH6.2 and then incubated with rabbit anti-nestin antibody at 4.0 µg/ml for 60 minutes. The reaction was developed using MACH 1 universal HRP polymer detection system and visualized with 3'3-diamino-benzidine substrate (DAB).

Immunohistochemistry with Anti-Nestin antibody. Tissue: mouse kidney (Left) and human kidney (Right). Fixation: formalin-fixed, paraffin-embedded tissue. Antigen retrieval: heat-induced. Primary antibody: 5 µg/ml. Staining: antibody as precipitated red signal with a hematoxylin purple nuclear counterstain.

Western blot using Biorbyt's Affinity Purified anti-Nestin antibody shows detection of a band ~220 kDa corresponding to human Nestin (arrowhead). Undifferentiated HCN-1A human brain cortical neuron neuronal progenitor lysate (lane 1), or differentiated HCN-1A human brain cortical neuron neuronal progenitor lysate (lane 2) were separated by SDS-PAGE using 4-20% gradient gel. After transfer onto nitrocellulose, the membrane was blocked and then probed with the primary antibody diluted to 1:2000 overnight at 4°C. The membrane was then washed and reacted with a 1:10000 dilution of peroxidase conjugated affinity purified Gt-a-Rabbit IgG [H&L] MX (orb347673) for 45 min at room temperature.

Western blot using Biorbyt's Affinity Purified anti-Nestin antibody shows detection of a band ~220 kDa corresponding to mouse Nestin (arrowhead). Approximately 30 g of MEF whole cell lysate was separated by SDS-PAGE using a 4-20% gradient gel. After transfer onto nitrocellulose, the membrane was blocked and then probed with the primary antibody diluted to 1:2000 overnight at 4°C. The membrane was then washed and reacted with a 1:10000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature.