Human/Mouse/Rat Bcl-2 Antibody Summary
Ala2-Asp211
Accession # P10415
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
Detection of Human, Mouse, and Rat Bcl‑2 by Western Blot. Western blot shows lysates of THP-1 human acute monocytic leukemia cell line, KG-1 human acute myelogenous leukemia cell line, CTLL-2 mouse cytotoxic T cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat Bcl-2 Monoclonal Antibody (Catalog # MAB8272) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Bcl-2 at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Bcl‑2 in Human Lymphoma. Bcl-2 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Mouse Anti-Human/Mouse/Rat Bcl-2 Monoclonal Antibody (Catalog # MAB8272) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Western Blot Shows Human Bcl‑2 Specificity by Using Knockout Cell Line. Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Bcl-2 knockout HeLa cell line (KO). PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat Bcl-2 Monoclonal Antibody (Catalog # MAB8272) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Bcl-2 at approximately 24 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Bcl-2 by Western Blot Cholesterol reversed metformin-mediated anticancer potential.Cells were treated with metformin (2 mM) alone and in combination with metformin (2 mM) plus cholesterol (10 μg/ml). (A) Cell viability (MTT) assay was performed to check the reverse effect of cholesterol on metformin- inhibited cell viability. Bars are representing the relative cell viability after 24 hr of treatment. Values represent mean ± SEM of triplicate measurements, **p <.001 vs. control and *p <.001 vs. metformin. (B) Scratch assay was performed. MDA-MB-231 cell monolayers were scratched and incubated with metformin (2 mM) alone and in combination with metformin (2 mM) plus cholesterol (10 μg/ml). The cell monolayers were photographed, and represented photos at 0 hr and 24 hr for control, metformin and metformin plus cholesterol cells were shown here. (C) The unfilled gap areas between two ends of the scratch were measured and subsequently plotted. Values represent mean ± SEM of triplicate measurements, **p <.01 vs. control at 0 hr and **p <.01 vs. metformin at 24 hr. (D) Colony formation assay was performed. After 24 hr of cell seeding, cells were treated with metformin (1mM) alone and in combination with metformin (1mM) plus cholesterol (5 μg/ml). After 5 days, formed colonies were stained with crystal violet. Photos of the wells were taken by a camera. (E) Soft agar assay of MDA-MB-231 cells in presence and absence of metformin (1mM) alone and in combination with metformin (1mM) plus cholesterol (5 μg/ml). Day 9 pictures were shown in the figure. Arrows depict the spheres. (F, G and H) RT-PCR analysis was performed using total RNA isolated from metformin, metformin (1mM) plus cholesterol treated and untreated cells, and gene specific primers. Cholesterol treatment reverses the metformin inhibited genes expressions (Bcl-xL [F], Zeb1 [G] and BMI1 [H]) (compare lane 2 vs. lane 3). (I and J) Western blot analysis was performed using total proteins from metformin, metformin (1mM) plus cholesterol treated and untreated cells, and specific antibodies. Cholesterol treatment reverses the metformin inhibited genes expression (BCL2 [I] and vimentin [J]) (compare lane 2 vs. lane 3). (K) Gelatin zymography was performed using equal amount of conditioned medium of metformin, metformin (2 mM) plus cholesterol (10 μg/ml) treated and untreated cells. Metformin-inhibited MMP activity was increased by treatment of cholesterol (compare lane 2 vs. lane 3). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30625181), licensed under a CC-BY license. Not internally tested by R&D Systems.
Reconstitution Calculator
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Bcl-2
Bcl-2 is a member of a family of proteins that regulates outer mitochondrial membrane permeability (1, 2). Bcl-2 is an anti-apoptotic member that prevents release of cytochrome c from the mitochondria intermembrane space into the cytosol. Bcl-2 is present on the outer mitochondrial membrane and is also found on other membranes in some cell types. Natural Bcl-2 contains a carboxyl-terminal mitochondria targeting sequence. Recombinant Bcl-2, missing the mitochondrial targeting sequence, maintains its ability to neutralize pro-apoptotic Bcl-2 family members. Neutralization by Bcl-2 appears to be through binding the BH3 region of pro-apoptotic Bcl-2 family members. This activity does not require the mitochondrial targeting sequence.
Product Datasheets
Citations for Human/Mouse/Rat Bcl-2 Antibody
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
12
Citations: Showing 1 - 10
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DL-3-n-butylphthalide protects endothelial cells against advanced glycation end product-induced injury by attenuating oxidative stress and inflammation responses
Authors: Chang-Yun Liu, Zhen-Hua Zhao, Zhi-Ting Chen, Chun-Hui Che, Zhang-Yu Zou, Xiao-Min Wu et al.
Experimental and Therapeutic Medicine
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Metformin induces pyroptosis in leptin receptor-defective hepatocytes via overactivation of the AMPK axis
Authors: B Liu, J Xu, L Lu, L Gao, S Zhu, Y Sui, T Cao, T Yang
Cell Death & Disease, 2023-02-03;14(2):82.
Species: Rat
Sample Types: Cell Lysates
Applications: Western Blot -
Human Umbilical Cord Mesenchymal Stem Cells Attenuate Severe Burn-Induced Multiple Organ Injury via Potentiating IGF-1 and BCL-2/BAX Pathway
Authors: H Wang, T Ba, Q Wang, L Yang, C Li, X Hao, Y Yin, L Liu
Stem Cells International, 2022-12-23;2022(0):5474289.
Species: Rat
Sample Types: Tissue Lysates
Applications: Western Blot -
Cyclo(Val-Pro) and Cyclo(Leu-Hydroxy-Pro) from Pseudomonas sp. (ABS-36) alleviates acute and chronic renal injury under in vitro and in vivo models (Ischemic reperfusion and unilateral ureter obstruction)
Authors: K Hira, P Sharma, A Mahale, O Prakash Ku, A Sajeli Beg
International immunopharmacology, 2021-12-29;103(0):108494.
Species: Rat
Sample Types: Cell Lysates
Applications: Western Blot -
Curcumin alleviates arsenic-induced toxicity in PC12�cells via modulating autophagy/apoptosis
Authors: MS Rahaman, S Banik, M Akter, MM Rahman, MT Sikder, T Hosokawa, T Saito, M Kurasaki
Ecotoxicol. Environ. Saf., 2020-05-25;200(0):110756.
Species: Rat
Sample Types: Whole Cell
Applications: Cell Culture -
Metformin exhibited anticancer activity by lowering cellular cholesterol content in breast cancer cells
Authors: A Sharma, S Bandyopadh, K Chowdhury, T Sharma, R Maheshwari, A Das, G Chakrabart, V Kumar, CC Mandal
PLoS ONE, 2019-01-09;14(1):e0209435.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Dexmedetomidine protects the developing rat brain against the neurotoxicity wrought by sevoflurane: role of autophagy and Drp1-Bax signaling
Authors: Y Shan, S Sun, F Yang, N Shang, H Liu
Drug Des Devel Ther, 2018-10-30;12(0):3617-3624.
Species: Rat
Sample Types: Tissue Homogenates
Applications: Western Blot -
FK506 Attenuates the MRP1-Mediated Chemoresistant Phenotype in Glioblastoma Stem-Like Cells
Authors: Á Torres, V Arriagada, JI Erices, MLÁ Toro, JD Rocha, I Niechi, C Carrasco, C Oyarzún, C Quezada
Int J Mol Sci, 2018-09-11;19(9):.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Exosomes derived from human umbilical cord mesenchymal stem cells improve myocardial repair via upregulation of Smad7
Authors: XL Wang, YY Zhao, L Sun, Y Shi, ZQ Li, XD Zhao, CG Xu, HG Ji, M Wang, WR Xu, W Zhu
Int. J. Mol. Med., 2018-02-16;0(0):.
Species: Human, Rat
Sample Types: Cell Lysates, Tissue Homogenates
Applications: Western Blot -
Hepatoprotective effect of calculus bovis sativus on nonalcoholic fatty liver disease in mice by inhibiting oxidative stress and apoptosis of hepatocytes
Authors: W He, Y Xu, C Zhang, J Lu, J Li, D Xiang, J Yang, M Chang, D Liu
Drug Des Devel Ther, 2017-12-05;11(0):3449-3460.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: Western Blot -
Icariin Protects Bone Marrow Mesenchymal Stem Cells Against Iron Overload Induced Dysfunction Through Mitochondrial Fusion and Fission, PI3K/AKT/mTOR and MAPK Pathways
Authors: Xudong Yao, Xingzhi Jing, Jiachao Guo, Kai Sun, Yi Deng, Yong Zhang et al.
Frontiers in Pharmacology
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Methamphetamine differentially affects BDNF and cell death factors in anatomically defined regions of the hippocampus
Authors: Melissa H. Galinato, Laura Orio, Chitra D. Mandyam
Neuroscience
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