Western blot analysis was used to detect protein expression in the ischemic tissue, as described by us in previous work21 (link),22 (link). Upon completion of electrophoresis, proteins were transferred to polyvinylidene fluoride membranes, which were incubated with primary antibodies (rabbit polyclonal anti-Bcl-2, rabbit polyclonal anti-BAX antibody, rabbit polyclonal anti-TN-Fα antibody, or rabbit polyclonal anti- IL-1β antibody; Abcam, Cambridge, MA, USA) for 24 h at 4°C. Next, membranes were washed with PBS three times for 6 min per wash, and then re-incubated with a secondary antibody (goat anti-rabbit IgG, Santa Cruz, Dallas, TX, USA; goat anti-mouse IgG, Abcam) for 1 h at room temperature. Finally, an enhanced chemiluminescence system was used to detect immunoreactive bands. Western blot images for each antibody, including β-actin, were analyzed using an image analysis program (Image J 1.42, NIH, Bethesda, MD, USA) to represent protein expression in terms of relative image density; mean protein expression from the control group was assigned a value of 1 to serve as a reference.
Rabbit polyclonal anti bax antibody
Manufactured by Abcam
Sourced in United States
Rabbit polyclonal anti-Bax antibody is a laboratory reagent. It is an antibody raised in rabbits that specifically binds to the Bax protein.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit polyclonal anti tnfα antibody, by Abcam (1 mentions)
Rabbit polyclonal anti il 1β antibody, by Abcam (1 mentions)
Goat anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Polyclonal rabbit anti bcl 2, by Abcam (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Sodium dodecyl sulfate polyacrylamide gel electrophoresis sds page gel preparation kit, by Beyotime (1 mentions)
Rabbit polyclonal anti bcl 2 antibody, by Abcam (1 mentions)
Polyvinylidene difluoride pvdf membranes, by Solarbio (1 mentions)
Goat anti rabbit igg horseradish peroxidase hrp, by Abcam (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Solarbio (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
3 protocols using rabbit polyclonal anti bax antibody
1
Quantifying Protein Expression in Ischemic Tissue
2
Detecting Bax Protein in Brain Tissue
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In order to detect Bax, a protein associated with apoptosis, the researchers employed the streptavidin-peroxidase method (Hassanen et al. 2022 (link)). The brain sections were first deparaffinized and hydrated, followed by three washes with phosphate-buffered saline (PBS) and the use of 3% H2O2 to neutralize the natural peroxidase activity. After another rinse with PBS, a blocking buffer consisting of normal low-lethal serum was applied. The sections were then subjected to immunohistochemical staining. They were incubated overnight at 4 ◦C with a diluted solution (1:50) of rabbit polyclonal anti-Bax antibody obtained from Abcam Co, USA. Next, the sections were treated with a secondary antibody and avidin–biotin complex. The stain 3,3′-diaminobenzidine was utilized as a chromogen, and Mayer's hematoxylin was used to counterstain the sections. Finally, the sections were examined under a light microscope.
3
Chitosan-based Cytotoxicity Evaluation
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Chitosan (DD > 90%) was purchased from Qingdao Biotemed Biomaterial Co., Ltd. (Qingdao, China). Chitosanase was prepared, purified, and identified in our laboratory. Fetal bovine serum (FBS) and RPMI 1640 medium were obtained from Gibco®, Life Technologies (Carlsbad, CA, USA). Calcein-AM was purchased from Fanbo Biochemical Co., Ltd. (Beijing, China). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and propidium iodide (PI) were obtained from Sigma–Aldrich (St. Louis, MO, USA). The RIPA lysis buffer and the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel preparation kit were obtained from Beyotime Biotechnology Co., Ltd. (Shanghai, China). The 3,3-diaminobenzidine (DAB) reagent kit, mouse tumor necrosis factor α (TNF-α) enzyme-linked immune sorbent assay (ELISA) kit, Mouse interleukin 2 (IL-2) ELISA Kit, Mouse Fas ELISA Kit and the Mouse Fas-L ELISA Kit were purchased from Boster Biological Engineering Co., Ltd. (Wuhan China). The rabbit polyclonal anti-Bcl-2 antibody, rabbit polyclonal anti-Bax antibody and goat anti-rabbit IgG Horse Radish Peroxidase (HRP) were purchased from Abcam Inc (Cambridge, MA, USA). Polyvinylidene difluoride (PVDF) membranes and bovine serum albumin (BSA) were obtained from Solarbio Science and Technology Co., Ltd. (Beijing, China).
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