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98322
Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit II
Primary Antibodies
Antibody Sampler Kit
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Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit II #98322

Citations (1)
Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells treated with cytochrome c (250 μM, 45 min) using BID Antibody #2002. The virtual lane view (left) shows two target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunoprecipitation of RL cell lysate using Bim (C34C5) Rabbit mAb (lane 1) or Rabbit (DA1E) mAb IgG XP® Isotype Control (lane 2). Western blot was performed using Bim (C34C5) Rabbit mAb. Protein A (HRP Conjugate) #12291 was used for secondary detection.
Simple Western™ analysis of lysates (1.0 mg/mL) from Raji cells using Bim (C34C5) Rabbit mAb #2933. The virtual lane view (left) shows three target bands (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Simple Western™ analysis of lysates (1 mg/mL) from HepG2 cells using Bax (D2E11) Rabbit mAb #5023. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Simple Western™ analysis of lysates (1 mg/mL) from OVCAR8 cells using Bad (D24A9) Rabbit mAb #9239. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from various cell lines using Bak (D4E4) Rabbit mAb.
Flow cytometric analysis of OVCAR8 cells using Bak (D4E4) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Western blot analysis of extracts from A549 cells, untreated (-) or treated with Doxorubicin #5927 (500 nM, overnight; +), using Puma (D30C10) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from KARPAS-299, RL, and L-540 cells lines using Noxa (D8L7U) Rabbit mAb. Cell Line Source: Dr. Abraham Karpas at the University of Cambridge.
Western blot analysis of extracts from Jurkat cells, untreated or etoposide-treated (25 µM), and RS4;11 cells, untreated or okadaic acid-treated (1 µM), using BID Antibody.
Western blot analysis of extracts from Raji, A20 and RL cells using Bim (C34C5) Rabbit mAb.
Western blot analysis of extracts from control HeLa cells (lane 1) or Bax knockout HeLa cells (lane 2) using Bax (D2E11) Rabbit mAb #5023 (upper), or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Bax-knockout HeLa cells confirms specificity of the antibody for Bax.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from various cell lines using Bok (D7V2N) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Bak (D4E4) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Puma (D30C10) Rabbit mAb.
Western blot analysis of extracts from serum starved Jurkat cells, untreated (-) or treated with TPA #4174 (10 ng/ml, 3 hr; +), using Noxa (D8L7U) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Bim siRNA I #6461 or SignalSilence® Bim siRNA II (+), using Bim (C34C5) Rabbit mAb #2933 and α-Tubulin (11H10) Rabbit mAb #2125. Bim (C34C5) Rabbit mAb confirms silencing of Bim expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Bim siRNA.
Western blot analysis of extracts from Raji and Ramos cell lines using Bik Antibody.
Western blot analysis of extracts from various cell lines using Bax (D2E11) Rabbit mAb. Brimmel et al. demonstated that Jurkat cells lack Bax protein expression (10).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Bok (hBok-Myc/DDK; +) using Bok (D7V2N) Rabbit mAb (upper) or Myc-Tag (71D10) Rabbit mAb #2278 (lower).
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Bak (D4E4) Rabbit mAb.
Immunoprecipitation of Puma from RL cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Puma (D30C10) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Puma (D30C10) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human Noxa protein (hNoxa; +), using Noxa (D8L7U) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Bim (C34C5) Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemical analysis of paraffin-embedded human breast caricnoma using Bax (D2E11) Rabbit mAb in the presence of control peptide (upper) or antigen-specific peptide (lower).
Immunohistochemical analysis of paraffin-embedded mouse prostate using Bak (D4E4) Rabbit mAb.
Immunoprecipitation of Noxa protein from KARPAS-299 cells using Rabbit (DA1E) mAb XP® Isotype Control #3900 (lane 2) or Noxa (D8L7U) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Noxa (D8L7U) Rabbit mAb. Cell Line Source: Dr. Abraham Karpas at the University of Cambridge.
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Bim (C34C5) Rabbit mAb performed on the Leica® BOND™ Rx.
Immunoprecipitation of Bax from HepG2 cells. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Bax (D2E11) Rabbit mAb, #5023. Western blot was performed using Bax (2D2) Mouse mAb, #89477, to prevent heavy and light chain IgG masking.
Confocal immunofluorescent analysis of OVCAR8 cells using Bak (D4E4) Rabbit mAb (green; left) showing colocalization with mitochondria that were labeled with MitoTracker® Red CMXRos (red; center). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Bim (C34C5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lymphoma using Bim (C34C5) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Bim (C34C5) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using Bim (C34C5) Rabbit mAb.
Confocal immunofluorescent analysis of MCF-7 cells using Bim Antibody (green) showing colocalization with mitochondria that have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of Raji cells using Bim (C34C5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
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To Purchase # 98322
Cat. # Size Qty. Price Inventory
98322T		 1 Kit  (9 x 20 microliters)

Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Bax (D2E11) Rabbit mAb 5023 20 µl
  • WB
  • IP
  • IHC
 H 20 Rabbit IgG
Bak (D4E4) Rabbit mAb 12105 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R Mk 25 Rabbit IgG
Bok (D7V2N) Rabbit mAb 86875 20 µl
  • WB
 H 22 Rabbit IgG
Bim (C34C5) Rabbit mAb 2933 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R 12, 15, 23 Rabbit IgG
Bad (D24A9) Rabbit mAb 9239 20 µl
  • WB
 H M R Mk 23 Rabbit IgG
BID Antibody 2002 20 µl
  • WB
  • IP
 H 15, 22 Rabbit 
Puma (D30C10) Rabbit mAb 12450 20 µl
  • WB
  • IP
 H 23 Rabbit IgG
Noxa (D8L7U) Rabbit mAb 14766 20 µl
  • WB
  • IP
 H 10 Rabbit IgG
Bik Antibody 4592 20 µl
  • WB
 H 20 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Rab Goat 

Product Description

The Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit II provides an economical means to examine several members of the Bcl-2 family. The kit contains enough primary antibody to perform two western blot experiments.

Specificity / Sensitivity

Each antibody in the Pro-Apoptosis Bcl-2 Family Antibody Sampler Kit II recognizes endogenous levels of its specific target. Bim (C34C5) Rabbit mAb detects endogenous levels of total Bim (EL, L, and S isoforms) protein. BID Antibody (Human Specific) detects endogenous levels of both the full length (22 kDa) and cleaved large fragment (15 kDa) of human BID. Puma (D30C10) Rabbit mAb cross-reacts with a protein of unknown origin at 60 kDa. Noxa (D8L7U) Rabbit mAb cross-reacts with multiple unidentified proteins, most notably at 35, 50, and 80 kDa.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu45 of human Bax, Gly75 of human Bak, Val88 of human Bok, Pro25 of human Bim, Pro102 of human Bad, Leu45 of human Bax, Pro25 of human Bim, residues near the carboxy terminus of human Puma, and residues near the amino terminus of human Noxa. Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the amino-terminus of human Bik or residues surrounding the cleavage site of human BID. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

Background

The Bcl-2 family consists of a number of evolutionarily conserved proteins containing Bcl-2 homology domains (BH) that regulate apoptosis through control of mitochondrial membrane permeability and release of cytochrome c (1-3). Four BH domains have been identified (BH1-4) that mediate protein interactions. The family can be separated into three groups based upon function and sequence homology: pro-survival members include Bcl-2, Bcl-xL, Mcl-1, A1 and Bcl-w; pro-apoptotic proteins include Bax, Bak and Bok; and "BH3 only" proteins Bad, Bik, Bid, Puma, Bim, Bmf, Noxa and Hrk. Interactions between death-promoting and death-suppressing Bcl-2 family members has led to a rheostat model in which the ratio of pro-apoptotic and anti-apoptotic proteins controls cell fate (4). Thus, pro-survival members exert their behavior by binding to and antagonizing death-promoting members. In general, the "BH3-only members" can bind to and antagonize the pro-survival proteins leading to increased apoptosis (5). While some redundancy of this system likely exists, tissue specificity, transcriptional and post-translational regulation of many of these family members can account for distinct physiological roles.

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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