Antibodies to bcl-2 Protein Family - Product Review 04

Introduction

Bcl-2 and related cytoplasmic proteins are key regulators of apoptosis, the cell suicide program critical for development, tissue homeostasis, and protection against pathogens.

Those most similar to Bcl-2 promote cell survival by inhibiting adapters needed for activation of the proteases (caspases) that dismantle the cell. More distant relatives instead promote apoptosis, apparently through mechanisms that include displacing the adapters from the pro-survival proteins. Thus, for many but not all apoptotic signals, the balance between these competing activities determines cell fate. Bcl-2 family members are essential for maintenance of major organ systems, and mutations affecting them are implicated in cancer.

At least 15 Bcl-2 family members have been identified in mammalian cells and several others in viruses. All members possess at least one of four conserved motifs known as Bcl-2 homology domains (BH1 to BH4). Most pro-survival members, which can inhibit apoptosis in the face of a wide variety of cytotoxic insults, contain at least BH1 and BH2, and those most similar to Bcl-2 have all four BH domains. The two pro-apoptotic subfamilies differ markedly in their relatedness to Bcl-2. Bax, Bak, and Bok (also called Mtd), which contain BH1, BH2, and BH3, resemble Bcl-2 fairly closely. In contrast, the seven other known mammalian "killers" possess only the central short (9 to 16 residue) BH3 domain; they are otherwise unrelated to any known protein, and only Bik and Blk are similar to each other. These "BH3 domain" proteins may well represent the physiological antagonists of the pro-survival proteins.

The Bcl2 family members are also characterized by dimerizing to further modulate apoptosis. Bag1, for example, has been found to form a heterodimer with Bcl2 resulting in the enhancement of the anti-apoptotic effect of Bcl2. Bax and Bak have been shown to play a critical role in cytochrome c release from mitochondria and thus initiate apoptosis. Bax exerts a pro-apoptotic rather than an anti-apoptotic effect on cells. Bax targets mitochondrial membranes, inducing mitochondrial damage and cell death in a caspase-independent manner. Bad plays a critical role in the Bax-mediated apoptosis pathway by dimerizing with BclxL, causing the displacment of Bax. The displacement of Bax allows apoptosis to proceed. BclxS, a shorter version of BclxL (lacking amino acids 126-188), apparently utilizes a different pathway than Bax to induce cell death. Some research suggests that BclxS uses a novel mechanism for regulating caspase or it may use an alternate cell death effector pathway.





Antibody Tools for Detection of bcl-2 Family Members

Antibody products for detection of many of the above listed bcl-2 family members are available at Acris Antibodies. All antibodies are thoroughly tested for different immunological detection methods. For more informations like host, isotype, species cross-reactivity etc. see table 1 below. For pricing info and datasheet download use the links provided.



Figure 1: Staining of human tonsil using bcl-2 antibody DM130. Note cytoplasmic and memebrane staining of follicular cells. No staining of reactive cells in the germinal center

Figure 1: Staining of human tonsil using bcl-2 antibody DM130. Note cytoplasmic and memebrane staining of follicular cells. No staining of reactive cells in the germinal center

Figure 2:  Staining of Hodgkin lymphoma using bcl-x antibody AM00285PU-N. Note staining of RS cells

Figure 2: Staining of Hodgkin lymphoma using bcl-x antibody AM00285PU-N. Note staining of RS cells

Figure 3:  Staining of Hodgkin lymphoma using bcl-x antibody AP00535PU-N. Note staining of RS cells.

Figure 3: Staining of Hodgkin lymphoma using bcl-x antibody AP00535PU-N. Note staining of RS cells.

Figure 4: Staining of human tonsil using Mcl-1 antibody AM00277PU-N. Note nuclear staining of cells.

Figure 4: Staining of human tonsil using Mcl-1 antibody AM00277PU-N. Note nuclear staining of cells.

Figure 5:  Staining of breast cancer using Bag-1 antibody DM335. Note nuclear and cytoplasmic staining of tumor cells

Figure 5: Staining of breast cancer using Bag-1 antibody DM335. Note nuclear and cytoplasmic staining of tumor cells

Figure 6: Staining of Hodgkin lymphoma using bax antibody DM246. Note membrane staining of RS cells

Figure 6: Staining of Hodgkin lymphoma using bax antibody DM246. Note membrane staining of RS cells

Figure 7: Staining of human tonsil using Bak antibody AP00539PU-N. Note cytoplasmic staining of mucosal epithelial cells.

Figure 7: Staining of human tonsil using Bak antibody AP00539PU-N. Note cytoplasmic staining of mucosal epithelial cells.

Figure 8:  Western blot detection of human bim using SP7027P in K562 (left) and A549 cell lysates (right)

Figure 8: Western blot detection of human bim using SP7027P in K562 (left) and A549 cell lysates (right)

Figure 9: Western blot detection of human bmf using SP1414P in HepG2 (left) and 293 cell lysates (right)

Figure 9: Western blot detection of human bmf using SP1414P in HepG2 (left) and 293 cell lysates (right)

Available antibody panel to bcl-2 Protein Family

Anti- bcl-2, bcl-x, Bcl-xL , Mcl-1, Bag-1, bax, bak, Blk, Bim, Bad, Bmf, NOXA, PUMA-alpha, PUMA beta*



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Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  
AM11017PU-N

Bcl-2-like 8 pSer155/118 antibody

Figure 1. Western analysis of cell extracts from 293 cells transfected with Flag-Bad, using Phospho-mBad-Ser155 Antibody (Cat #AM11017PU-N).
Mouse IgG1 27AT381 Purified Hu, Ms E, WB
0.4 ml / €340.00
    Acris Antibodies GmbH
AM11018PU-N

Bcl-2-like 8 pSer155/118 antibody

Bcl-2-like 8 Mouse IgG1 27AT937 Purified Hu, Ms E, WB
0.4 ml / €340.00
    Acris Antibodies GmbH
MAB0113

Bcl-2-like 8 pSer155/118 antibody

Western analysis of cell extracts from 293 cells transfected with Flag-Bad, using Bad (phospho S155) monoclonal antibody, clone 27AT381 ( Cat # MAB0113 ). Mouse IgG1 27AT381 Hu, Ms E, WB
0.4 ml / €350.00
    Abnova Taiwan Corp.

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