Bcl-2 Antibody (YA590S)
(Synonyms: Apoptosis regulator Bcl-2, BCL2)
Based on 1 Customer Validation
Bcl-2 Antibody (YA590S) is a Mouse-derived and non-conjugated IgG2a monoclonal antibody, targeting to Bcl-2.
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Host:
Mouse
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Isotype:
IgG
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Application:
WB, IHC-P, FC
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Reactivity :
Human, Mouse
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Formulation:
Supplied in PBS (pH7.4), 40% Glycerol, 0.05% BSA, 0.05% Sodium Azide.
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Conjugation:
Non-conjugated
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Applications
| Application |
WB
WB: Western Blot
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IHC-P
IHC-P: Immunohistochemistry-Paraffin
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FC
FC: Flow Cytometry
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| Dilution Ratio | 1:1000-1:4000 | 1:500-1:1500 | 1:800 |
Product Details
Bcl-2 Antibody (YA590S) is a Mouse-derived and non-conjugated IgG2a monoclonal antibody, targeting to Bcl-2.
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Host Mouse
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Clonality Monoclonal
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Species ReactivityHuman, Mouse
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Observed Molecular WeightObserved band size: 26 kDa
Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
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Calculated Molecular Weight Predicted band size: 26 kDa
Recombinant protein of human Bcl-2 aa1-211 (HY-P7537)
Endogenous
Affinity purified
Non-conjugated
Unmodified
IgG
Product Properties
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Appearance
Solution
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Formulation
Supplied in PBS (pH7.4), 40% Glycerol, 0.05% BSA, 0.05% Sodium Azide.
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Concentration
Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration
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Storage & Stability
Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
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Shipping
Shipping with blue ice.
Verification Images
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Western blot analysis of Whole cell lysate from RL (+) (lane 2, 5 μg), THP-1 (+) (lane 3, 20 μg), Jurkat (+) (lane 4, 20 μg), and HT-29(-) (lane 5, 20 μg) using Bcl-2 antibody(HY-P880009). Proteins were transferred to a PVDF membrane and blocked with non-fat milk in TBST at 4°C overnight. The primary antibody (1/2000) and Loading control antibody (β-Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature. -
Western blot analysis of Whole cell lysate from RL (+) (lane 2, 5 μg), THP-1 (+) (lane 3, 20 μg), Jurkat (+) (lane 4, 20 μg), and HT-29 (-) (lane 5, 20 μg) using Bcl-2 antibody(HY-P880009). Proteins were transferred to a PVDF membrane and blocked with non-fat milk in TBST at 4°C overnight. The primary antibody (1/4000) and Loading control antibody(β-Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature. -
Western blot analysis of lysate from Mouse spleen (lane 2, 40 μg) and Rat spleen (lane 3, 40 ug) using Bcl-2 antibody(HY-P880009). The primary antibody (1/1000) and Loading control antibody (β-Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1hour at room temperature. -
Western blot analysis of extracts from RAW.264.7 (lane 2, 40 μg) and Mouse spleen (lane 3, 40 ug) using Bcl-2 antibody(HY-P880009). The primary antibody (1/1000) and Loading control antibody (β-Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 30 minutes at room temperature. -
Western blot analysis of Whole cell lysate from RL (+) (lane 2, 5 μg), THP-1 (+) (lane 3, 20 μg), Jurkat (+) (lane 4, 20 μg), and HT-29(-) (lane 5, 20 μg) using Bcl-2 antibody(HY-P880009). Proteins were transferred to a PVDF membrane and blocked with non-fat milk in TBST at 4°C overnight. The primary antibody (1/2000) , competitor's antibody (1/1000) and Loading control antibody (β-Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature. -
Western blot analysis was performed on protein extracts (25 μg) from Jurkat (lane 2), THP-1 (lane 3), and MCF-7 (lane 4) using Bcl-2 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot® Turbo™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST (HY-K1025) for 1 h at room temperature. The primary antibody (1: 2000) and loading control antibody β-actin Antibody (HRP) (HY-P80993) (1:10000) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibody (HY-P8001/HY-P8004) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit (HY-K1005). -
Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue using Bcl-2 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 8.0) for 1.5 minutes. The tissues were blocked in 3% H2O2 for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P880009, 1/1500) for overnight at 4°C temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded Human Thyroid cancer tissue using Bcl-2 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 8.0) for 1.5 minutes. The tissues were blocked in 3% H2O2 for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P880009, 1/1500) for overnight at 4°C temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded Human Prostate cancer tissue using Bcl-2 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 8.0) for 1.5 minutes. The tissues were blocked in 3% H2O2 for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P880009, 1/1500) for overnight at 4°C temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded Human Breast cancer tissue using Bcl-2 antibody. The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 8.0) for 1.5 minutes. The tissues were blocked in 3% H2O2 for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P880009, 1/1500) for overnight at 4°C temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of 1X105 RL cells (+) ( red) and 1X105 HT_x001e_29 cells (-) ( blue) labeling Bcl-2 Antibody (HY-P880009). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then stained with the primary antibody at 1/800 dilution (1 μg/mL) for an hour at 4°C. AF488-conjugated Goat Anti-Rabbit IgG H&L (HY-P8005) was used as the secondary antibody at 1/1000 dilution for 30 minutes at 4°C. Rabbit IgG Isotype Control was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (dotted line).
Background
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Function
Bcl-2 (B-cell lymphoma 2) is a central anti-apoptotic regulator that preserves mitochondrial integrity by preventing mitochondrial outer membrane permeabilization (MOMP), a critical checkpoint controlling cytochrome c release and caspase activation during intrinsic apoptosis[1][2]. Mechanistically, Bcl-2 functions through direct interactions with pro-apoptotic BCL-2 family proteins, including Bax and Bak, thereby suppressing their oligomerization and limiting mitochondrial membrane disruption[3][4]. The protein also integrates signals transmitted by BH3-only proteins, which regulate the balance between cell survival and programmed cell death under physiological and pathological stress conditions[3][5]. Dysregulated Bcl-2 expression contributes to apoptosis resistance in multiple malignancies and supports tumor cell survival by maintaining mitochondrial homeostasis despite cellular stress[6][7]. In disease models, the anti-apoptotic activity of Bcl-2 is closely linked to mitochondrial apoptosis pathways that represent major therapeutic targets in hematologic and solid cancers[6][8]. Compared with related anti-apoptotic isoforms such as Mcl-1 and Bcl-B, Bcl-2 displays broader interactions with both Bax and Bak, whereas Mcl-1 and Bcl-B show preferential regulation of Bak- and Bax-dependent apoptotic pathways, respectively[4]. For experimental applications, BH3-mimetic inhibitors disrupt the interaction between Bcl-2 and pro-apoptotic partners, thereby restoring mitochondrial apoptosis and providing a widely used strategy for mechanistic studies and targeted cancer research[7][8].
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Subcellular Localization
Mitochondrion outer membrane; Single-pass membrane protein; Nucleus membrane; Single-pass membrane protein; Endoplasmic reticulum membrane; Single-pass membrane protein; Cytoplasm
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Expression
Tissue_specificity:Expression in multiple organizations -
Isoforms & Post-Translational Modification
P10415 has 2 isomers: P10415-1: 26266 Da (predicted); P10415-2: 22337 Da (predicted).
Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity (PubMed:11368354). Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle (PubMed:11368354). In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases (PubMed:11368354). Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, which stimulates starvation-induced autophagy (PubMed:10567572, PubMed:18570871). Dephosphorylated by protein phosphatase 2A (PP2A) (By similarity);Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity;Monoubiquitinated by PRKN, leading to an increase in its stability (PubMed:20889974). Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome (PubMed:23431138). Ubiquitinated by XIAP, leading to its degradation by the proteasome (PubMed:29020630) -
Subunit
Forms homodimers, and heterodimers with BAX, BAD, BAK and Bcl-X(L). Heterodimerization with BAX requires intact BH1 and BH2 motifs, and is necessary for anti-apoptotic activity (PubMed:25609812, PubMed:8183370).
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SwissProt ID
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Synonyms
Apoptosis regulator Bcl-2, BCL2
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Research Field
Apoptosis
Documentation
References
[4]. Zhai D, et al. Differential regulation of Bax and Bak by anti-apoptotic Bcl-2 family proteins Bcl-B and Mcl-1. J Biol Chem. 2008 Apr 11;283(15):9580-6. [Content Brief]
[5]. Lomonosova E, et al. BH3-only proteins in apoptosis and beyond: an overview. Oncogene. 2008 Dec;27 Suppl 1(Suppl 1):S2-19. [Content Brief]
[8]. Kang MH, et al. Bcl-2 inhibitors: targeting mitochondrial apoptotic pathways in cancer therapy. Clin Cancer Res. 2009 Feb 15;15(4):1126-32. [Content Brief]
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