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.
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Key Product Details
Validated by
Species Reactivity
Validated:
Cited:
Applications
Validated:
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Label
Antibody Source
Product Specifications
Immunogen
Ala2-Asp211
Accession # P10415
Specificity
Clonality
Host
Isotype
Scientific Data Images for Bcl-2 Antibody (625509)
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 cholesteroDetection of hplqjjbtewfjidvyz6xc.jpg by Western Blot
Western blotting analysis for proteins in rat livers and BRL 3 A cells co-cultured with blood-derived exosomes from ZDF and Zucker lean rats.A Western blotting analysis for the livers of SD, Zucker lean and ZDF rats at 6 months post different treatments, (as indicated). B BRL 3 A cells were subject to Western blotting analysis at 48 h and 2 weeks post exosome co-culture. C Quantitative analysis of B. Except for AMPK (n = 10) and p-AMPK (n = 10), n = 12. Band intensities in B were quantified via Image Lab (Bio-Rad, Hercules, CA, USA) and the protein expression level was normalized to the level of the beta -actin. Data are presented on a logarithmic scale as the fold change in expression relative to controls (after coculture with exosomes extracted from 48 h HFD-fed Zucker lean rats in the absence of metformin; Zucker lean+HFD). *p < 0.05 vs. controls; #p < 0.05 vs. cultures after coculture with exosomes extracted from 48 h HFD-fed ZDF rats in the absence of metformin; ZDF + HFD; delta p < 0.05 vs. cultures with exosomes extracted from 2-week HFD-fed Zucker lean rats in the absence of metformin (Zucker lean + HFD); †p < 0.05 vs. cultures with exosomes extracted from 2-week HFD-fed ZDF rats in the absence of metformin (ZDF + HFD). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36737598), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Bcl-2 by Western Blot
Western blotting analysis for proteins in rat livers and BRL 3 A cells co-cultured with blood-derived exosomes from ZDF and Zucker lean rats.A Western blotting analysis for the livers of SD, Zucker lean and ZDF rats at 6 months post different treatments, (as indicated). B BRL 3 A cells were subject to Western blotting analysis at 48 h and 2 weeks post exosome co-culture. C Quantitative analysis of B. Except for AMPK (n = 10) and p-AMPK (n = 10), n = 12. Band intensities in B were quantified via Image Lab (Bio-Rad, Hercules, CA, USA) and the protein expression level was normalized to the level of the beta -actin. Data are presented on a logarithmic scale as the fold change in expression relative to controls (after coculture with exosomes extracted from 48 h HFD-fed Zucker lean rats in the absence of metformin; Zucker lean+HFD). *p < 0.05 vs. controls; #p < 0.05 vs. cultures after coculture with exosomes extracted from 48 h HFD-fed ZDF rats in the absence of metformin; ZDF + HFD; delta p < 0.05 vs. cultures with exosomes extracted from 2-week HFD-fed Zucker lean rats in the absence of metformin (Zucker lean + HFD); †p < 0.05 vs. cultures with exosomes extracted from 2-week HFD-fed ZDF rats in the absence of metformin (ZDF + HFD). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36737598), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Bcl-2 Antibody (625509)
Immunohistochemistry
Sample: Immersion fixed paraffin-embedded sections of human lymphoma
Knockout Validated
Western Blot
Sample: 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
Reviewed Applications
Read 1 review rated 5 using MAB8272 in the following applications:
Formulation, Preparation, and Storage
Purification
Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.
Formulation
Shipping
Stability & 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.
Calculators
Background: Bcl-2
References
- Gross, A. et al. (1999) Genes and Develop. 13:1899.
- Kroemer, G. (1997) Nature Med. 3:614.
Long Name
Alternate Names
Gene Symbol
UniProt
Additional Bcl-2 Products
Product Documents for Bcl-2 Antibody (625509)
Certificate of Analysis
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Product Specific Notices for Bcl-2 Antibody (625509)
For research use only
Related Research Areas
Citations for Bcl-2 Antibody (625509)
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Application: Immunohistochemistry-FrozenSample Tested: LiverSpecies: MouseVerified Customer | Posted 08/20/2021Bcl-2 Antibody, brown staining
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars