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34811
Apoptosis Antibody Sampler Kit II
Primary Antibodies
Antibody Sampler Kit
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Apoptosis Antibody Sampler Kit II #34811

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Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Cytochrome C using Caspase-3 (D3R6Y) Rabbit mAb #14220. The virtual lane view (left) shows the 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.
Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Etoposide (25 uM, 5 hours) using PARP (46D11) Rabbit mAb #9532. The virtual lane view (left) shows the 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.
Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and PARP (46D11) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the GET4 transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.
Simple Western™ analysis of lysates (0.1 mg/mL) from Jurkat cells treated with Cytochrome C using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb #9664. The virtual lane view (left) shows the 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.
Western blot analysis of extracts from THP-1 cells, untreated (-) or treated (+) as indicated with: Cycloheximide #2112 (10 μg/mL, 18 hr) followed by Human Tumor Necrosis Factor-α (hTNF-α) #8902 (20 ng/mL, 4 hr) using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb (upper), Caspase-8 (1C12) Mouse mAb #9746 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from Jurkat cells, untreated (-) or treated with Etoposide #2200 (1 μM, 18 hr; +), using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb (upper), Caspase-8 (1C12) Mouse mAb #9746 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunoprecipitation of cleaved caspase-8 protein from HeLa cell extracts. Cells were treated with Staurosporine #9953 (10 μM, 3 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb. Western blot analysis was performed using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Flow cytometric analysis of serum starved HCT 116 cells, untreated (blue) or treated with Staurosporine #9953 (1 μM, 4 hr; green), using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of HCT 116 cells (left) or CRISPR/Cas9 Casp8 knockout (KO) HCT 116 cells (right), both serum starved and then treated with Staurosporine #9953 (1 μM, 4 hr) using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Simple Western™ analysis of Jurkat cell lysates (1 mg/mL) treated with Cytochrome C (0.25mg/mL, 45 min) using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb #98134. The virtual lane view (left) shows the target band (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.
Western blot analysis of various cell lines, untreated (-) or treated with Staurosporine #9953 (1 μM; 3 hr) or with Etoposide #2200 (25 μM, overnight), using Caspase-3 (D3R6Y) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). MCF7 cells are negative for caspase-3 expression.
Western blot analysis of extracts from HeLa and Jurkat cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 3 hr; +) or Etoposide #2200 (25 μM, overnight; +), using Cleaved-Caspase-9 (Asp315) (D8I9E) Rabbit mAb (upper), Caspase-9 (C9) Mouse mAb #9508 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from serum starved HeLa or HT-29 cells, untreated (-) or treated with Staurosporine #9953 (1 μM; +), using Cleaved PARP (Asp214) (E2T4K) Mouse mAb (upper), total PARP (46D11) Rabbit mAb #9532 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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.
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 Jurkat cells (human), L929 cells (mouse), and C6 cells (rat), untreated or treated with staurosporine or cytochrome c as indicated, using Caspase 9 (C9) Mouse mAb.
Western blot analysis of extracts from control HEK293 cells (lane 1) or PARP knockout HEK293 cells (lane 2) using PARP (46D11) Rabbit mAb #9532 (upper) or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the PARP knockout HEK293 cells confirms the specificity of the antibody for PARP.
Western blot analysis of extracts from C6 (rat), NIH/3T3 (mouse), and Jurkat (human) cells, untreated or treated with staurosporine #9953 (1uM, 3hrs) or etoposide #2200 (25uM, 5hrs) as indicated, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.
Western blot analysis of extracts from control HeLa cells (lane 1) or Caspase-8 knockout HeLa cells (lane 2) using Caspase-8 (1C12) Mouse mAb #9746 (upper), or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Caspase-8-knockout HeLa cells confirms specificity of the antibody for Caspase-8.
Western blot analysis of extracts from HCT 116 and CRISPR/Cas9 caspase-8 knockout (KO) HCT 116 cells, untreated (-) or treated with Staurosporine #9953 (1 μM, 4 hr; +), using Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb (upper), Caspase-8 (1C12) Mouse mAb #9746 (middle), or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from HCT116 cells (lane 1) or CASP3 knock-out cells (lane 2) using Caspase-3 (D3R6Y) Rabbit mAb #14220 (upper), and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The absence of signal in the CASP3 knock-out HCT116 cells confirms specificity of the antibody for CASP3.
Confocal immunofluorescent analysis of HeLa cells, serum-starved (left) or treated with Staurosporine #9953 (1 μM, 3 hr; right), using Cleaved Caspase-9 (Asp315) (D8I9E) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunoprecipitation of cleaved PARP (Asp214) from serum-starved HeLa cells treated with Staurosporine #9953 (1 μM, 3 hr). Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Cleaved PARP (Asp214) (E2T4K) Mouse mAb. Western blot was performed using Cleaved PARP (Asp214) (E2T4K) Mouse mAb. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.
Western blot analysis of extracts from THP-1 cells, untreated or treated with TNF-α and cycloheximide as well as control extracts from SW620 and A20 cell lines, using PARP (46D11) Rabbit mAb.
Immunoprecipitation of extracts from Jurkat cells, untreated or etoposide-treated (25uM, 5hrs), using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb. Western blot was performed using the same antibody.
Western blot analysis of extracts from HCT 116 and HCT 116 Caspase-8 knockout cells, untreated (-) or treated with staurosporine #9953 (1 μM, 4 hr; +), using #9746 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The absence of signal in the HCT 116 knockout cells confirms the specificity of the antibody for Caspase-8.
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Etoposide #2200 (25 μM, 18 hr; green) using Cleaved Caspase-9 (Asp315) (D8I9E) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffin-embedded HeLa cell pellet, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Western blot analysis of extracts from SKW6.4 cells, untreated or anti-Fas-treated (1 µg/ml), and Jurkat cells, untreated or etoposide-treated (25 µM), using Caspase-8 (1C12) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemical analysis of paraffin-embedded mouse embryo, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb in the presence of control peptide (left) or Cleaved Caspase-3 (Asp175) Blocking Peptide (#1050) (right).
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemical analysis using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb on SignalSlide® Cleaved Caspase-3 IHC Controls #8104 (paraffin-embedded Jurkat cells, untreated (left) or etoposide-treated (right)).
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.
Confocal immunofluorescent analysis of serum-starved HeLa cells, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved PARP (Asp214) (E2T4K) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with etoposide (25 μM, 18 hr; green), using Cleaved PARP1 (Asp214) (E2T4K) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415. Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Confocal immunofluorescent images of HT-29 cells, untreated (left) or Staurosporine #9953 treated (right) labeled with Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with etoposide #2200 (green), using Cleaved Caspase-3(Asp175) (5A1E) Rabbit mAb compared to a nonspecific negative control antibody (red).
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To Purchase # 34811
Cat. # Size Qty. Price
34811T		 1 Kit  (8 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Caspase-3 (D3R6Y) Rabbit mAb 14220 20 µl
  • WB
  • IP
 H M R Mk 35, 19, 17 Rabbit IgG
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 9664 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R Mk 17, 19 Rabbit IgG
PARP (46D11) Rabbit mAb 9532 20 µl
  • WB
  • IP
  • eCLIP
 H M R Mk 116, 89 Rabbit 
Cleaved-PARP (Asp214) (E2T4K) Mouse mAb 32563 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H 89 Mouse IgG1
Caspase-9 (C9) Mouse mAb 9508 20 µl
  • WB
 H M R Hm Mk 47/37/35 (H). 49/39/37 (M). 51/40/38 (R). Mouse IgG1
Cleaved Caspase-9 (Asp315) (D8I9E) Rabbit mAb 20750 20 µl
  • WB
  • IP
  • IF
  • F
 H 35 Rabbit IgG
Caspase-8 (1C12) Mouse mAb 9746 20 µl
  • WB
  • IP
 H 18, 43, 57 Mouse IgG1
Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb 98134 20 µl
  • WB
  • IP
  • IF
  • F
 H 18, 41, 43 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
  • WB
 Horse 

Product Description

The Apoptosis Antibody Sampler Kit II provides an economical means of detecting selected targets involved in apoptosis. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Background

Apoptosis is a regulated physiological process leading to cell death. Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Initiator caspases (including 8, 9, 10, and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6, and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF, and lamin A and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase (1). Proapoptotic stimuli include FasL, TNF-α, DNA damage and ER stress. Fas and TNFR activate caspase-8 and -10 (2), DNA damage leads to the activation of caspase-9 and ER stress leads to the calcium-mediated activation of caspase-12 (3). The inhibitor of apoptosis protein (IAP) family includes XIAP and survivin and functions by binding and inhibiting several caspases (4,5). Smac/Diablo, a mitochondrial protein, is released into the cytosol upon mitochondrial stress and competes with caspases for binding of IAPs. The interaction of Smac/Diablo with IAPs relieves the inhibitory effects of IAPs on caspases (6).

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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