Bcl-2 Antibody (100/D5)

BCL2 was detected in immersion fixed paraffin-embedded sections of human Non-Hodgkin’s Lymphoma using Mouse Anti-Human BCL2, Monoclonal Antibody (Catalog #NBP2-15200) at a 1:500 dilution at 37 ° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). TTissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the membrane. Protocol available in COMET™ Panel Builder.

Western Blot: Bcl-2 Antibody (100/D5) [NBP2-15200] - Western Blot Analysis of human MCF-7 cell lysate using Bcl-2 Antibody (100/D5).

Immunohistochemistry-Paraffin: Bcl-2 Antibody (100/D5) [NBP2-15200] - Formalin-paraffin non-Hodgkin's lymphoma stained with Bcl-2 Ab (100/D5). Note nuclear membrane & cytoplasmic staining.

Flow Cytometry: Bcl-2 Antibody (100/D5) [NBP2-15200] - Intracellular staining of Bcl-2 in fixed and permeabilized Jurkat cells using Bcl-2 antibody (red) and isotype control (green) at 0.5 ug/10^6 cells.

Western Blot: Bcl-2 Antibody (100/D5) [NBP2-15200] - analysis of Bcl-2 in human skin using Bcl-2 antibody at 2 ug/ml.

• Reactivity: Hu, Mu(-), Rt(-)
• Applications: WB, Flow, ICC/IF, IHC, IP
• Clone: 100/D5
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

Bcl-2 Antibody (100/D5) Summary

• Description: 200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP2-33315)
  
  Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80 C.
• Immunogen: A synthetic peptide, aa41-54 (GAAPAPGIFSSQPG-Cys) of human Bcl-2 protein. (Uniprot: P10415 )
• Localization: Outer mitochondrial membranes and endoplasmic reticulum as well as nuclear membranes.
• Marker: Apoptosis & Follicular Lymphoma Marker
• Specificity: This antibody recognizes a protein of 25-26kDa, identified as the Bcl-2 alpha oncoprotein. It shows no cross-reaction with Bcl-x or Bax protein. Expression of Bcl-2 alpha oncoprotein inhibits the programmed cell death (apoptosis). In most follicular lymphomas, neoplastic germinal centers express high levels of Bcl-2 alpha protein, whereas the normal or hyperplastic germinal centers are negative. Consequently, this antibody is valuable when distinguishing between reactive and neoplastic follicular proliferation in lymph node biopsies. It may also be used in distinguishing between those follicular lymphomas that express Bcl-2 protein and the small number in which the neoplastic cells are Bcl-2 negative.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: BCL2
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow (Intracellular)
  • Flow Cytometry 1-2 ug/million cells
  • Immunocytochemistry/ Immunofluorescence 1-2 ug/ml
  • Immunohistochemistry
  • Immunohistochemistry-Frozen 0.5-1ug/ml
  • Immunohistochemistry-Paraffin 1-2 ug/ml
  • Immunoprecipitation
  • Western Blot 1-2 ug/ml
• Application Notes: Use in IP reported in scientific literature (PMID:35304128) Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
  Optimal dilution for a specific application should be determined..

Reactivity Notes

Does not react with Mouse or Rat.

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein A or G purified

Alternate Names for Bcl-2 Antibody (100/D5)

• apoptosis regulator Bcl-2
• B-cell CLL/lymphoma 2
• Bcl2
• Bcl-2

Background

Bcl-2 (Bcl2) is the founding member of the Bcl-2 family. Family members have pivotal roles in regulating apoptosis or death signaling pathways through their control of mitochondrial permeability and cytochrome release (reviewed in Anvekar, 2011; Martinou and Youle, 2011). Bcl-2 derives its name from B cell lymphoma 2 where was first found to be highly expressed in follicular lymphomas with 14;18 reciprocal translocations. There are two isoforms, alpha and beta, generated by alternative splicing and differing in their carboxy termini. Human Bcl-2 alpha is a 239 amino acid (aa) protein and human Bcl-2 beta is a 205 aa protein. The 100/D5 antibody [also known as clone Bcl-2/100 (Kren, 2004, Kaur, 2004)] recognizes both Bcl-2 isoforms (Pezzela et al, 1990).
Bcl-2 is over expressed in neoplastic germinal centers of a majority of follicular lymphomas, whereas the normal or hyperplastic germinal centers are primarily negative for Bcl-2 expression. Upregulation has also been described in a number of other types of tumors. Bcl-2 expression is often considered to be a marker of cell death status, and over or high expression has often been tied to anti-apoptotic states, or resistance to death. However, the actual status of vulnerability to death can depend on the balance of other Bcl-2 family members present, their interaction with one another, as well as other factors.
This antibody has been extensively used to monitor the status of Bcl-2 expression in a myriad of normal and cancerous tissue types, developmental stages, as well as in cultured cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Bcl-2 Antibody (NBP2-15200)(11)

Publications using NBP2-15200

• Tian D, You X, Ye J et al. BMSCs overexpressing hBcl2 can resist myelin-induced apoptosis and promote repair after spinal cord injury in rats Research Square 2023-08-28 (IHC, ICC/IF, WB, Rat, Mouse)
  
  Details:
  WB dilution 1:100; ICC/If dilution 1:100; IHC dilution 1:100
• Sevinc SK, Karadeniz M, Sen A et al. Apoptotic and antiproliferative effects of Urtica dioica L. extract on K562 chronic myeloid leukemia cell line Indian Journal of Experimental Biology 2023-11-01 (WB)
• Mandic M, Misirkic Marjanovic M, Vucicevic L et al. MAP kinase-dependent autophagy controls phorbol myristate acetate-induced macrophage differentiation of HL-60 leukemia cells Life sciences 2022-05-15 [PMID: 35304128] (IP, Human)
• Mega Tiber P, Kocyigit Sevinc S, Kilinc O, Orun O Biological effects of whole Z.Officinale extract on chronic myeloid leukemia cell line K562. Gene. 2019-04-15 [PMID: 30684525] (WB, Human)
  
  Details:
  Citation using the Azide free version of this antibody.
• Pradhan N, Pratheek BM, Garai A et al. Induction of apoptosis by Fe(salen)Cl through caspase-dependent pathway specifically in tumor cells. Cell Biol. Int. 2014-05-07 [PMID: 24804954] (Flow Cytometry Control, Human)
  
  Details:
  Mouse splenocytes, human PBMC, Jurkat, EL4: Fig 5. GADPH was detected at ~36 kDa.
• Moschovi M, Koultouki E, Stefanaki K et al. Prognostic significance of angiogenesis in relation to Ki-67, p-53, p-27, and bcl-2 expression in embryonal tumors. Pediatr Neurosurg. 2011-01-01 [PMID: 22309998] (IHC-P)
  
  Details:
  IHC (paraffin): Embryonal tumors, Table 1.
• Tralongo V, Becchina G, Nagar C et al. Primary follicular lymphoma of the epididymis positive for t(14;18)(q32;q21)/IGH-BCL2 and negative for BCL2 protein expression: a case report. J Med Case Rep. 2012-01-18 [PMID: 22257663] (IHC-P)
• Kren L, Brazdil J, Hermanova M et al. Prognostic significance of anti-apoptosis proteins survivin and bcl-2 in non-small cell lung carcinomas: a clinicopathologic study of 102 cases. Appl Immunohistochem Mol Morphol. 2004-03-01 [PMID: 15163019] (IHC-P)
  
  Details:
  IHC (paraffin): lung cancer.
• Kaur P, Kallakury BS, Sheehan CE et al. Survivin and Bcl-2 expression in prostatic adenocarcinomas. Arch Pathol Lab Med. 2004-01-01 [PMID: 14692814] (IHC-P)
  
  Details:
  IHC (paraffin): Prostate adenocarcinoma, Figs 2,3.
• Pezzella F, Tse AG, Cordell JL et al. Expression of the bcl-2 oncogene protein is not specific for the 14;18 chromosomal translocation. Am J Pathol. 1990-08-01 [PMID: 2201196] (ICC/IF, IHC-Fr, WB, IHC-P)
  
  Details:
  WB: Fig 1 (normal human spleen), the Bcl-2 alpha isoform was detected in both reducing and non-reducing conditions. IHC (frozen)/ IHC (paraffin)/ IF/ICC: Various human cell types and tissues: Figs 2,7 and Tables 1-3. See publication for specific staining
• Kocyigit Sevinc S, Karadeniz M, Sen A et al. Biological Effects of Urtica Dioica Extract on K562 Chronic Myeloid Leukemia Cell Line SSRN Electronic Journal 2022-10-29 (WB, Human)

Bioinformatics

• Gene Symbol: BCL2
• Uniprot:
  • P10415()
  • P10415()
  • P10415()