Rabbit anti bcl 2 antibody

Manufactured by Proteintech

Sourced in United States, China

The Rabbit anti-Bcl-2 antibody is a laboratory reagent designed for the detection and analysis of the Bcl-2 protein. Bcl-2 is a key regulator of apoptosis, or programmed cell death. This antibody can be used in various immunoassay techniques to study the expression and localization of Bcl-2 in biological samples.

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5 protocols using rabbit anti bcl 2 antibody

Western Blotting Analysis of Liver Proteins

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Liver samples were processed from −80°C storage and Western blotting performed as described [20 (link)]. The primary antibodies used are as follows: KLF2 (1 : 400, rabbit anti-KLF2 antibody, Biosynthesis Biotechnology, Beijing, China), phosphorylated eNOS at Ser1177 (1 : 1000, rabbit anti-phosphorylated eNOS antibody, Cell Signaling, Danvers, MA), total eNOS (1 : 1000, rabbit anti-eNOS antibody, Cell Signaling, Danvers, MA), Bax (1 : 1000, rabbit anti-Bax antibody, Proteintech, Manchester, UK), and Bcl-2 (1 : 1000, rabbit anti-Bcl-2 antibody, Proteintech, Manchester, UK). The bands were revealed by chemiluminescence ECL reagent (Proteintech, Manchester, UK), and protein expression was quantified by densitometric analysis using the Quantity One software package (Hertfordshire, UK). All bands were assayed for β-actin (1 : 1000, rabbit anti-β-actin antibody, Proteintech, Manchester, UK) content as standardization of sample loading.

Liu Z., Zhang X., Xiao Q., Ye S., Lai C.H., Luo J., Huang X., Wang W., Zeng C., Zhong Z., Fan X., Xia Z., Xiong Y., Mao X., Ye Q, & Wang Y. (2017). Pretreatment Donors after Circulatory Death with Simvastatin Alleviates Liver Ischemia Reperfusion Injury through a KLF2-Dependent Mechanism in Rat. Oxidative Medicine and Cellular Longevity, 2017, 3861914.

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Western Blot Analysis of Inflammasome Proteins

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Lysates were obtained by lysing the cell monolayer with phenylmethanesulfonyl fluoride (PMSF) and radioimmunoprecipitation assay (RIPA) lysis buffer (1:100). Total protein was separated in 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and then transferred to polyvinylidene difluoride (PVDF) membranes. Proteins were detected after labeling with specific primary antibodies [rabbit anti-NLRP3 (1:500, Bioss, China), rabbit anti-ASC polyclonal antibody (1:500, Bioss, China), rabbit anti-Caspase-1 (1:1000, Abcam, USA), rabbit anti cleaved Caspase1 (Arg317)/P10 (1:1000, Affinity, USA), rabbit anti-IL-1β antibody (1:1000, Abcam, USA), rabbit anti-BAX antibody (1:10000, Proteintech, USA), and rabbit anti-Bcl2 antibody (1:1500, Proteintech, USA)] followed by HRP-coupled secondary antibodies. The signals were detected by the addition of the chemiluminescent substrate (ECL Western Blot Kit, CWBIO, China). The protein amount was determined using Image J software. The level of GAPDH (1:1500, Proteintech, USA) was used to normalize the signal intensities.

Huang Y., Sun X., Juan Z., Zhang R., Wang R., Meng S., Zhou J., Li Y., Xu K, & Xie K. (2021). Dexmedetomidine attenuates myocardial ischemia-reperfusion injury in vitro by inhibiting NLRP3 Inflammasome activation. BMC Anesthesiology, 21, 104.

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Immunohistochemical Analysis of Cardiac Markers

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Paraffin-embedded cardiac tissues were cut into 5-μm-thick cross-sections and
deparaffinized prior to staining using a standard protocol. Immunohistochemical
staining was performed according to the manufacturer's instructions (Zsbio,
Beijing, China) using antibodies against Bax (rabbit anti-Bax antibody, 1:200;
Proteintech, Wuhan, China), Bcl-2 (rabbit anti-Bcl-2 antibody, 1:200;
Proteintech), NRF2 (rabbit anti-NRF2 antibody, 1:200; Proteintech), NOX4 (rabbit
anti-NOX4 antibody, 1:200; Proteintech), and HO-1 (rabbit anti-HO-1 antibody,
1:200; SOLARBIO, Beijing, China). All sections were examined using a BX40
upright light microscope.

Guo W., Long X., Lv M., Deng S., Liu D, & Yang Q. (2022). Effect of thymoquinone on sepsis-induced cardiac damage via anti-inflammatory and anti-apoptotic mechanisms. The Journal of International Medical Research, 50(9), 03000605221118680.

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Immunohistochemical Analysis of Cardiac Markers

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For immunohistochemical staining, the heart sections were deparaffinized and rehydrated. Next, the sections were blocked with 3% H₂O₂ in methanol for 15 min to inactivate the endogenous peroxidases and incubated overnight at 4°C with the primary antibodies: CD36 (rabbit anti-CD36 antibody, 1 : 200; Proteintech, Wuhan, China), SCD-1 (rabbit anti-SCD-1 antibody, 1 : 200; Abcam, England), PGC1-ɑ (rabbit anti-PGC1-ɑ antibody, 1 : 200; Proteintech), CPT-1 (rabbit anti-CPT-1 antibody, 1 : 200; Proteintech), p-AMPK (rabbit anti-p-AMPK antibody, 1 : 100; Proteintech), PPAR-α (rabbit anti-PPAR-α antibody, 1 : 50; Proteintech), BAX (rabbit anti-BAX antibody, 1 : 200; Proteintech), Bcl-2 (rabbit anti-Bcl-2 antibody, 1 : 200; Proteintech), and cleaved caspase-3 (rabbit anti-cleaved caspase-3 antibody, 1 : 400; Cell Signaling Technology, USA). The sections were incubated with goat anti-rabbit HRP secondary antibody (Histofine Simple Stain Kit; Nichirei, Tokyo, Japan) for 30 min at room temperature. All sections were examined using an Olympus light microscope (Olympus, Tokyo, Japan).

Pei Z., Wang X., Yang C., Dong M, & Wang F. (2021). Recombinant Human Growth Hormone Inhibits Lipotoxicity, Oxidative Stress, and Apoptosis in a Mouse Model of Diabetic Cardiomyopathy. Oxidative Medicine and Cellular Longevity, 2021, 3899356.

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Western Blot Analysis of Cardiac Fibrosis Markers

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The cardiac tissues were harvested, and protein extracts prepared, according to established methods. Extracts were separated in sodium dodecyl sulphate–polyacrylamide electrophoresis gels (8–15%) and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford, MA, USA). The membranes were blocked with 5% milk and then incubated with indicated primary antibodies at 4 °C overnight. Primary antibodies against TGF-β (rabbit anti-TGF-β antibody, 1 : 1000; Proteintech, Wuhan, China), Smad3 (rabbit anti-Smad antibody, 1 : 1000; Proteintech), collagen I (rabbit anti-collagen I antibody, 1 : 1000; Proteintech), collagen III (rabbit anti-collagen III antibody, 1 : 1000; Proteintech), α-SMA (rabbit anti-α-SMA antibody, 1 : 1000; Proteintech), P53 (rabbit anti-P53 antibody, 1 : 1000; Proteintech), bcl-2(rabbit anti-bcl-2 antibody, 1 : 1000; Proteintech), anti-β-actin (1 : 1000; Cell Signaling Technology). After washing, the membranes were incubated with the appropriate secondary antibody (anti-rabbit Ig-G, 1 : 1000; Cell Signaling Technology) for 1 hour. This analysis was carried out independently three times. Protein levels were expressed as protein/β-actin ratios to minimise loading differences. The relative signal intensity was quantified using NIH ImageJ software.

Pei Z., Hu J., Bai Q., Liu B., Cheng D., Liu H., Na R, & Yu Q. (2018). Thymoquinone protects against cardiac damage from doxorubicin-induced heart failure in Sprague-Dawley rats. RSC Advances, 8(26), 14633-14639.

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