Western blotting for the determination of Bax, Bcl-2, BDNF, TrkB was conducted as previously described method (Park et al., 2019 (link); Song et al., 2018 (link)). The hippocampus tissues were homogenized on ice and lysed in a lysis buffer containing 50 mM Tris-HCl (pH, 7.5), 150 mM NaCl, 0.5% deoxycholic acid, 1% Nonidet P40, 0.1% sodium dodecyl sulfate (SDS), 1 mM phenylmethylsulfonyl fluoride, and 100-mg/mL leupeptin. Protein content was measured using a Bio-Rad colorimetric protein assay kit (Bio-Rad, Hercules, CA, USA). Protein of 30 μg was separated on SDS-polyacrylamide gels and transferred onto a nitrocellulose membrane, which was incubated with mouse β-actin antibody (1:1,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse Bax (1:1,000; Santa Cruz Biotechnology), Bcl-2 (1:1,000; Santa Cruz Biotechnology), and rabbit BDNF (1:1,000; Santa Cruz Bio-technology), TrkB (1:1,000; Santa Cruz Biotechnology). Horseradish peroxidase-conjugated anti-mouse for β-actin, Bax, Bcl-2 and anti-rabbit for BDNF, TrkB were used as secondary antibodies.
Rabbit bdnf
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Rabbit BDNF is a laboratory reagent used for research purposes. It is a recombinant protein derived from the brain-derived neurotrophic factor (BDNF) gene of the rabbit species. BDNF is a growth factor that plays a crucial role in the survival and differentiation of neurons.
Automatically generated - may contain errors
Lab products found in correlation
Colorimetric protein assay kit, by Bio-Rad (2 mentions)
Mouse β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
P akt, by Santa Cruz Biotechnology (1 mentions)
P pdk, by Cell Signaling Technology (1 mentions)
Mouse β actin, by Santa Cruz Biotechnology (1 mentions)
P pi3k, by Santa Cruz Biotechnology (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
2 protocols using rabbit bdnf
1
Hippocampal Western Blot Analysis
2
Western Blot Analysis of Hippocampal Proteins
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Hippocampal tissue was homogenized on ice and lysed in a radioimmunoprecipitation (RIPA) cell lysis buffer (1X) with ethylenediaminetetraacetic acid (EDTA). Protein content was measured using a Bio-Rad colorimetric protein assay kit (Bio-Rad, Hercules). We separated 20 µg of protein on SDS-polyacrylamide gels and transferred the protein onto a nitrocellulose blotting membrane, which was incubated with the following primary antibodies: mouse β-actin (1:1000; Santa Cruz Biotech), rabbit BDNF (Santa Cruz Biotech, sc-20981), TrkB (1:1000; Santa Cruz Biotech, sc-119), IR (1:1000; cell signaling, #3095), p-IR (Tyr1162/1163) (1:1000; cell application, CB 486), PI3K (Santa Cruz Biotech, sc-1637), p-PI3K (Tyr467, sc-293115) (Santa Cruz Biotech, sc-293115) PDK (cell signaling, #3062), p-PDK (Ser241) (1:1000; cell signaling, #3061), AKT (Santa Cruz Biotech, sc-8312), p-AKT (Ser 473) (Santa Cruz Biotech, sc-135651), GSK-3β (cell signaling, #9315), and p-GSK3β (Ser 9) (1:1000; cell signaling, #9323). Horseradish peroxidase-conjugated anti-rabbit secondary antibody was used for BDNF, TrkB, IR, p-IR, PDK, p-PI3K, p-PDK, AKT, p-AKT, GSK-3β, and p-GSK-3β and anti-mouse secondary antibody was used for PI3K and β-actin. Experiments were performed under normal lab conditions and at room temperature (except for the transferred membrane).
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