Primary antibodies recognizing pathological changes in tau (Alz-50, conformation; and CP13, phospho-Serine 202; generous gift from Dr. Peter Davies, Albert Einstein College of Medicine) were used for IHC and WB. Anti-β-tubulin (Cat. # ab5392, Abcam) and anti-MAP2 (Cat. #ab56676, Abcam) primary antibodies were used as controls in WB and IHC, respectively. The anti-tubulin antibody was used to standardize protein levels; and for lack of availability of a suitable commercial rat-specific pan-tau antibody that provides consistent staining of tau. Horseradish peroxidase (HRP)-labeled secondary antibodies (Southern Biotech, Birmingham, AL, USA) were used for WB, and fluorescence-labeled secondary antibodies (Molecular Probes, Invitrogen) were used for IHC.
Horseradish peroxidase hrp labeled secondary antibodies
Manufactured by Southern Biotech
Sourced in United States
Horseradish peroxidase (HRP)-labeled secondary antibodies are laboratory reagents used for immunodetection applications. They consist of secondary antibodies conjugated with the enzyme horseradish peroxidase. These antibodies can be used to detect and quantify target proteins in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence labeled secondary antibody, by Thermo Fisher Scientific (1 mentions)
Ab5392, by Abcam (1 mentions)
Ab56676, by Abcam (1 mentions)
Sds page gel, by Thermo Fisher Scientific (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Anti β actin, by US Biological (1 mentions)
Anti vegf, by Santa Cruz Biotechnology (1 mentions)
2 protocols using horseradish peroxidase hrp labeled secondary antibodies
1
Immunodetection of Pathological Tau
2
Western Blot Analysis of VEGF and EMP2
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Cell lysates were prepared and lysed in Laemmli buffer. Proteins were separated on 4% to 20% SDS-PAGE gel (Thermo Fisher Scientific) under reducing conditions, transferred to nitrocellulose membranes (GE Healthcare, Chicago, IL, USA), and blocked with 10% nonfat dry milk in Tris-buffered saline with 0.1% Tween 20. Membranes were probed with the following primary antibodies: anti-VEGF (1 μg/mL; Santa Cruz Biotechnology, Dallas, TX, USA); anti-EMP2 (1:2000; polyclonal antibody; created in our lab and previously published26 (link)); anti-β-actin (0.5 μg/mL; US Biological, Salem, MA, USA). Protein bands were detected using horseradish peroxidase (HRP)-labeled secondary antibodies (Southern Biotechnology, Birmingham, AL, USA), followed by chemiluminescence (ECL; EMD Millipore, Burlington, MA, USA). Densitometric analysis of the protein bands were performed using NIH ImageJ software. Loading variability between samples were normalized to β-actin loading controls. At least three independent experiments were performed, and statistical significance was evaluated using Student's t-test.
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