Primary antibodies against Bcl-2, Bax, Bad, and Bcl-xL were procured from Abcam (USA). Akt, p-Akt, mTORC1, p-mTORC1, mTORC2, p-mTORC2, phosphatase and tensin homolog (PTEN), glycogen synthase kinase 3β (GSK-3β), p- GSK-3β (Ser9), NF-κBp65, and cleaved caspase-3 were procured from Cell Signaling Technology (Danvers, MA, USA). CyclinD1, β-actin, and horseradish- and peroxidase-labeled IgG secondary antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA) and used for expression analysis. Cell lysis buffer for Western blotting analysis was purchased from Beyotime Institute of Biotechnology (Beijing, China). Mangiferin and isoflurane were procured from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals used for analysis were obtained from Sigma-Aldrich.
Primary antibodies against bcl 2
Manufactured by Abcam
Sourced in United States
Primary antibodies against Bcl-2. Bcl-2 is an anti-apoptotic protein that plays a crucial role in the regulation of programmed cell death. These primary antibodies can be used to detect and quantify Bcl-2 expression in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Mangiferin, by Merck Group (1 mentions)
Isoflurane, by Merck Group (1 mentions)
Mtorc1, by Cell Signaling Technology (1 mentions)
Bcl xl, by Abcam (1 mentions)
Phosphatase and tensin homolog pten, by Cell Signaling Technology (1 mentions)
P mtorc1, by Cell Signaling Technology (1 mentions)
Mtorc2, by Cell Signaling Technology (1 mentions)
P mtorc2, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Antibody against cleaved caspase3, by Affinity Biosciences (1 mentions)
Alexa fluor 488 labeled, by Abcam (1 mentions)
Collagenase type 2, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
C myc, by Abcam (1 mentions)
4 protocols using primary antibodies against bcl 2
1
Molecular Signaling Pathway Analysis
2
Western Blot Analysis of Apoptosis Regulators
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The expression changes of Bcl-2 and Bax protein were determined using a western blot assay. After transfection, cells were collected and lysed with RIPA buffer to extract protein. The protein concentration was measured using the BCA protein assay kit (Beyotime). Then protein was separated using denaturing 10% SDS-PAGE and transferred to nitrocellulose membranes. The membranes were treated with blocking buffer and incubated with the primary antibodies against Bcl-2, Bax, or GAPDH (Abcam) at 4°C overnight, respectively. After washing with TBST, the membranes were incubated with the secondary antibody (Abcam) at 37°C for 1 h. Protein signals were detected using an ECL detection system and quantified using ImageJ software.
3
Investigating Cellular Stress Mechanisms
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tert-Butylhydroquinone (t-BHQ) and bafilomycin A1 were obtained from MCE (Monmouth Junction, NJ, USA). tert-Butyl hydroperoxide solution (TBHP), chloroquine (CQ), and type II collagenases were purchased from Sigma-Aldrich (St. Louis, MO, USA). Primary antibodies against BCL-2, Bax, SIRT3, NRF2, LC3, and collagen II were obtained from Abcam (Cambridge, MA, USA). Antibodies against GAPDH and p62 were acquired from Proteintech (Chicago, IL, USA). Antibodies against TOM20 and aggrecan were from Santa Cruz Biotechnology. The antibody against cleaved caspase-3 was from Affinity (Cincinnati, OH, USA). Alexa Fluor 488-labeled and Alexa Fluor 647-labeled goat anti-rabbit/rat secondary antibodies were purchased from Abcam (Cambridge, MA, USA). 4,6-Diamidino-2-phenylindole (DAPI) was obtained from Beyotime (Shanghai, China). Reagents for NPC culture were obtained from Gibco (Grand Island, NY, USA).
4
Diagnosis of DLBCL via IHC Scoring
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The diagnosis of DLBCL in all pathologic specimens was confirmed and reviewed by at least two hematopathologists with expertise at our institute. Tissue fixation and processing were performed using standard methods. Tissue microarrays that contained two representative 1-mm cores from each tumor were prepared using AutoTiss 1000 tissue microarrayer (EverBio Technology Inc., Taipei, Taiwan). IHC staining was performed as previously reported (25 (link)). Primary antibodies against BCL2 (Abcam, Cambridge, UK) and c-Myc (Abcam) were used at dilutions of 1/250 and 1/200, respectively. A high-sensitivity diaminobenzidine (DAB+) chromogenic substrate system, ab80436–EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC Kit (Abcam), was used for detection. All patient cases were stained and scored semiquantitatively in 10% increments by two observers (JL Liu and CE Huang) without knowledge of patient outcome or each other’s results of interpretation, and only lymphoma cells were scored. Previously established cutoff points used to define double-expressor lymphoma (≥50% BCL2-positive lymphoma cells and ≥40% MYC-positive lymphoma cells) (12 (link),13 (link)) were used to stratify tumors as high BCL2 expression and high MYC expression, respectively. Representative slides of various staining results on IHC studies for BCL2 and MYC expression are shown in Figures S1,S2
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