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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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

BAG1 (C-term) antibody

Western blot analysis in whole cell lysate from PC-3 human prostate cancer cells probed with Rabbit anti Human BAG1 (AP05585PU-N) at 1(A) and 2(B) µg/ml. Rabbit IgG Purified Hu, Ms, Rt WB
0.1 mg / €420.00
  Acris Antibodies GmbH

BAG1 (N-term) antibody

Immunohistochemical staining of human lymph node with Rabbit anti Human BAG1 (AP05586PU-N) Rabbit IgG Purified Hu, Ms WB
0.1 mg / €330.00
  Acris Antibodies GmbH

BAG1 antibody

Kidney: Formalin-Fixed Paraffin-Embedded (FFPE) Rabbit Aff - Purified Hu, Ms, Rt P, WB
50 µg / €470.00
  Acris Antibodies GmbH

BAG1 (N-term) antibody

Figure 1. Formalin-Fixed Paraffin-Embedded (FFPE) on Adrenal. Rabbit Purified Hu, Ms P, WB
50 µg / €470.00
  Acris Antibodies GmbH

BAG1 (C-term) antibody

Western blot analysis of BAG1 Antibody (C-term) (AP17147PU-N) in MDA-MB231 cell line lysates (35 µg/lane). BAG1 (arrow) was detected using the purified Pab. Rabbit Hu WB
0.4 ml / €370.00
  Acris Antibodies GmbH

BAG1 antibody

BAG1 MaxPab rabbit polyclonal antibody. Western Blot analysis of BAG1 expression in human placenta. Rabbit Purified Hu E(detection), WB
0.1 mg / €380.00
  Abnova Taiwan Corp.

BAG1 antibody

Formalin-fixed, paraffin-embedded sections of human colorectal cancer stained for Bag1. Using Bag1 polyclonal antibody ( Cat # PAB0327 ) at 1 : 2000. Hematoxylin-eosin counterstain. Samples from two patients ( A and B ) are shown. A1 and B1 are higher magnifications from A and B, respectively. Rabbit Bov, Hu, Ms, Rt IP, P
50 µl / €390.00
  Abnova Taiwan Corp.

BAG1 antibody

BAG1 Rabbit Hu, Ms, Rt C, WB
0.2 mg / €390.00
  Abnova Taiwan Corp.

BAG2 antibody

Western Blot analysis of BAG2 expression in transfected 293T cell line (H00009532-T01) by BAG2 MaxPab polyclonal antibody.

Lane 1: BAG2 transfected lysate(23.21 KDa).
Lane 2: Non-transfected lysate. Mouse Purified Hu E(detection), WB
50 µg / €370.00
  Abnova Taiwan Corp.

BAG2 antibody

Immunoprecipitation of BAG2 transfected lysate using anti-BAG2 MaxPab rabbit polyclonal antibody and Protein A Magnetic Bead (U0007), and immunoblotted with BAG2 MaxPab mouse polyclonal antibody (B01) (H00009532-B01). Rabbit Serum Hu E(detection), IP
0.1 ml / €380.00
  Abnova Taiwan Corp.

BAG2 antibody

Immunofluorescence confocal microscopy of BAG2. Using BAG2 polyclonal antibody ( Cat # PAB0329 ) at 1 : 2000. HeLa cells stably transfected with Myc-CHIP ( carboxyl terminus of Hsp70-interacting protein ) and growing on coverslips were left untreated ( A ) or heat shocked at 42 degrees for 30 min ( B ) . Cells were fixed with 3.7% paraformaldehyde, permeabilized with 0.5% Triton X-100 prior to antibody staining. BAG2 localized to the cytoplasm of untreated cells, and to the nucleus of cells subjected to heat shock. Please refer to Dai et al ( 2005 ) for additional information on HeLa cells stably transfected with Myc-CHIP. Rabbit Hu C, ICC/IF, IP, P, WB
50 µl / €400.00
  Abnova Taiwan Corp.

BAG2 antibody

BAG2 polyclonal antibody ( Cat # PAB6054 ) staining ( 0.3 µg/ml ) of HeLa lysate ( 35 µg protein in RIPA buffer ) . Primary incubation was 1 hour. Detected by chemiluminescence. Goat Hu E, WB
0.1 mg / €400.00
  Abnova Taiwan Corp.

BAG2 antibody

Human Kidney; Rabbit Anti-BAG2 Antibody. Catalog Number: ARP42177. Paraffin Embedded Tissue: Human Kidney. Cellular Data: Epithelial cells of renal tubule. Antibody Concentration: 4.0-8.0 ug/ml. Magnification: 400X; BAG2 antibody - C-terminal region (ARP42177_T100) in Human Kidney cells using Immunohistochemistry Rabbit Aff - Purified Bov, Can, Chk, Hu, Ms, Rt, Ze P, WB
0.1 mg / €310.00
  AVIVA Systems Biology

BAG2 antibody

Western Blot: BAG2 Antibody [NB100-1776] - NB100-1776 (0.3ug/ml) staining of Hela lysate (35ug protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence. Goat IgG Aff - Purified Hu E, WB
0.1 mg / €400.00
  Novus Biologicals Inc.

BAG3 antibody

Immunohistochemistry of formalin-fixed paraffin-embedded human pancreas tissue sections. Using BAG3 polyclonal antibody ( Cat # PAB0330 ) at 1 : 2000. A, normal pancreas. B and C, pancreatic cancer. Hematoxylin-eosin counterstain. Rabbit Hu, Ms C, IP, P, WB
50 µl / €470.00
  Abnova Taiwan Corp.

BAG4 antibody

Figure 1. Formalin-Fixed Paraffin-Embedded (FFPE) on Brain, cerebellum. Rabbit Purified Hu, Ms, Rt ICC/IF, P, WB
50 µg / €470.00
  Acris Antibodies GmbH

BAG4 (N-term) antibody

Immunocytochemical staining of Hela cells with Rabbit anti SODD (SP1301) Rabbit Aff - Purified Hu, Ms, Rt ICC/IF, WB
0.1 mg / €330.00
  Acris Antibodies GmbH

BAG4 (N-term) antibody

Immunocytochemical staining of Hela cells with Rabbit anti SODD (SP1301) Rabbit IgG Aff - Purified Hu, Ms, Rt ICC/IF, WB
50 µg / €280.00
  Acris Antibodies GmbH

BAG4 antibody

Immunoperoxidase of purified MaxPab antibody to BAG4 on formalin-fixed paraffin-embedded human stomach. [antibody concentration 3 ug/ml] Mouse Purified Hu E(detection), P, WB
50 µg / €370.00
  Abnova Taiwan Corp.

BAG4 antibody

Western Blot analysis of BAG4 expression in transfected 293T cell line (H00009530-T02) by BAG4 MaxPab polyclonal antibody.

Lane 1: BAG4 transfected lysate(49.60 KDa).
Lane 2: Non-transfected lysate. Rabbit Serum Hu E(detection), IP, WB
0.1 ml / €380.00
  Abnova Taiwan Corp.

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