Interested in promotions? | Click here >>
42867
Human Reactive Cell Death and Autophagy Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Human Reactive Cell Death and Autophagy Antibody Sampler Kit #42867

Reviews ()
Citations (0)
other Image 1 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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).

other Image 2 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Flow cytometric analysis of Jurkat cells, untreated (blue) or etoposide-treated (green), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.

other Image 3 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of HT-29 cells, untreated (-) or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), human TNF-α (hTNF-α, 20 ng/ml, 7 hr; +), SM-164 (100 nM, 7 hr; +), and necrostatin-1 (Nec-1, 50 μM, 7 hr; +), using Phospho-RIP (Ser166) (D1L3S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 4 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Confocal immunofluorescent analysis of HT-29 cells, untreated (left), pre-treated with Z-VAD (20 μM, 30 min) followed by treatment with SM-164 (100 nM) and Human Tumor Necrosis Factor-α (hTNF-α) #8902 (20 ng/mL, 6 hr; center), or pre-treated with Z-VAD followed by treatment with SM-164 and hTNF-α and post-processed with λ-phosphatase (right), using Phospho-RIPK3 (Ser227) (D6W2T) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

other Image 5 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of HT-29 cells, untreated (-), or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), human TNF-α (hTNF-α, 20 ng/ml, 7 hr; +), SM-164 (100 nM, 7 hr; +), and necrostatin-1 (Nec-1, 50 μM, 7 hr; +), using Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb (upper), or MLKL (D2I6N) Rabbit mAb #14993 (lower).

other Image 6 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.

other Image 7 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Confocal immunofluorescent analysis of THP-1 cells, differentiated with TPA #4174 (80 nM, 24 hr) and subsequently treated with (right) or without (left) Lipopolysaccharides (LPS) #14011 (1 μg/ml, 6 hr), using Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

other Image 8 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Confocal immunofluorescent analysis of MIA PaCa-2 (upper) and C2C12 (lower) cells, untreated (left), nutrient-starved with EBSS (2 hr; middle), or chloroquine treated (50 μM, 24 hr; right), using LC3B (E5Q2K) Mouse mAb (green) and β-Actin (D6A8) Rabbit mAb #8457 (red). Blue = Hoechst 33342 #4082 (fluorescent DNA dye).

other Image 9 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with chloroquine (50 μM overnight; right), using SQSTM1/p62 (D5L7G) Mouse mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

other Image 10 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from HeLa cells (lane 1) or SQSTM1 knock-out cells (lane 2) using SQSTM1/p62 (D5L7G) Mouse mAb #88588 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the SQSTM1 knock-out HeLa cells confirms specificity of the antibody for SQSTM1.

other Image 11 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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.

other Image 12 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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).

other Image 13 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (green). Actin filament were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

other Image 14 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of HT-29 cells, untreated (-) or treated with a combination of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), Human Tumor Necrosis Factor-α #8902 (hTNF-α, 20 ng/ml, 7 hr; +), and SM-164 (100 nM, 7 hr; +), using Phospho-RIP3 (Ser227) (D6W2T) Rabbit mAb. To confirm phospho-specificity, membranes were either untreated (left) or treated with Calf Intestinal Phosphatase (CIP; right).

other Image 15 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb in the presence of non-cleaved Gasdermin D peptide (left) or Asp275 cleavage-specific Gasdermin D peptide (right).

other Image 16 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunoprecipitation of Cleaved-IL-1β (Asp116) from extracts of THP-1 cells differentiated with TPA #4147 (80 nM, overnight) followed by treatment with Lipopolysaccharides (LPS) #14011 (1 μg/ml, 6 hr). Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is precipitated with Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb. Western blot was performed using Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb.

other Image 17 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using LC3B (E5Q2K) Mouse mAb.

other Image 18 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SQSTM1/p62 (D5L7G) Mouse mAb.

other Image 19 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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).

other Image 20 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human tonsil using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.

other Image 21 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of HT-29 cells, untreated (-) or treated with a combination of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), Human Tumor Necrosis Factor-α #8902 (hTNF-α, 20 ng/ml, 7 hr; +), SM-164 (100 nM, 7 hr; +), and necrostatin-1 (Nec-1, 50 μM, 7 hr; +), using Phospho-RIP3 (Ser227) (D6W2T) Rabbit mAb (upper), RIP3 (E1Z1D) Rabbit mAb #13526 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 22 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.

other Image 23 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of recombinant Human Interleukin-1β (hIL-1β) #8900 using Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb.

other Image 24 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using LC3B (E5Q2K) Mouse mAb.

other Image 25 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using SQSTM1/p62 (D5L7G) Mouse mAb.

other Image 26 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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)).

other Image 27 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr), Jurkat cells, untreated or etoposide-treated (25 μM, overnight), and THP-1 cells, untreated or cycloheximide-treated (CHX, 10 μg/ml, overnight) followed by treatment with TNF-α #8902 (20 ng/ml, 4 hr), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (upper), or total PARP Antibody #9542 (lower).

other Image 28 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of HT-29 cells or HT-29 RIPK1 KO cells, untreated (-) or treated with a combination of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), Human Tumor Necrosis Factor-α #8902 (hTNF-α, 20 ng/ml, 7 hr; +), and SM-164 (100 nM, 7 hr; +), using Phospho-RIP3 (Ser227) (D6W2T) Rabbit mAb (upper), RIP3 (E1Z1D) Rabbit mAb #13526 (middle) or β-Actin (D6A8) Rabbit mAb #8457 (lower). HT-29 RIPK1 KO cells were kindly provided by Dr. Junying Yuan, Harvard Medical School, Boston, MA.

other Image 29 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's Lymphoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.

other Image 30 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from cells or media collected from THP-1 cells, differentiated with TPA #4147 (80 nM, overnight) and subsequently treated with (+) or without (-) Lipopolysaccharides (LPS) #14011 (1 μg/ml, 6 hr), using Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb.

other Image 31 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using LC3B (E5Q2K) Mouse mAb.

other Image 32 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using SQSTM1/p62 (D5L7G) Mouse mAb.

other Image 33 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical staining of paraffin-embedded mouse embryo, showing cytoplasmic localization in apoptotic cells, using Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb.

other Image 34 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.

other Image 35 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using LC3B (E5Q2K) Mouse mAb.

other Image 36 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded HDLM-2 (left) and Daudi (right) cell pellets using SQSTM1/p62 (D5L7G) Mouse mAb.

other Image 37 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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.

other Image 38 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human spleen (left, positive) and skeletal muscle (right, negative) using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb.

other Image 39 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunoprecipitation of LC3B from HeLa cells treated with Chloroquine (50 μM, overnight) # 14774. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control, and lane 3 is LC3B (E5Q2K) Mouse mAb. Western blot was performed using LC3B (E5Q2K) Mouse mAb. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.

other Image 40 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunohistochemical analysis of paraffin-embedded human inflamed gall bladder using SQSTM1/p62 (D5L7G) Mouse mAb in the presence of control peptide (left) or antigen-specific peptide (right).

other Image 41 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

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.

other Image 42 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunoprecipitation of Cleaved Gasdermin D (Asp725) from THP-1 cells differentiated with TPA #4174 (50 ng/ml, overnight) and then treated with LPS #14011 (5 μg/ml, 6 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb. Western blot was performed using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.

other Image 43 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from HeLa, C2C12, and KNRK cells, untreated (-) or treated with Chloroquine (50 μM, overnight; +) #14774 using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 44 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Immunoprecipitation of SQSTM1/p62 protein from PANC-1 cell extracts. Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is SQSTM1/p62 (D5L7G) Mouse mAb. Western blot was performed using SQSTM1/p62 (D5L7G) Mouse mAb. A light chain-specific secondary antibody was used to avoid reactivity with heavy chain IgG.

other Image 45 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from THP-1 cells, differentiated with TPA #4174 (50 ng/ml, overnight) and then treated with LPS #14011 (5 μg/ml, indicated times), using Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb (upper), total Gasdermin D (L60) Antibody #93709 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 46 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from HeLa cells or HeLa cells with a knockout of LC3B (HeLa/LC3B KO) using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 47 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from A549 cells, untreated (-) or starved with Earle's Balanced Salt Solution (EBSS) (indicated times) using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb (lower).

other Image 48 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from various cell lines using SQSTM1/p62 (D5L7G) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 49 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from MCF7 and Raji cells, untreated (-) or treated with Torin 1 (250 nM, 5 hr; +) #14379, using LC3B (E5Q2K) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 50 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extract from HCT 116 cells, untreated (-) or starved with Earles Basic Salt Solution (EBSS; 4 hr; +) using SQSTM1/p62 (D5L7G) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 51 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot anlaysis of extracts from HeLa cells, untreated (-) or treated with Chloroquine #14774 (50 μM, overnight; +) using SQSTM1/p62 (D5L7G) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 52 - Human Reactive Cell Death and Autophagy Antibody Sampler Kit

Western blot analysis of extracts from HeLa cells transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® SQSTM1/p62 siRNA I #6394 (+), or SignalSilence® SQSTM1/p62 siRNA II #6399 (+), using SQSTM1/p62 (D5L7G) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

To Purchase # 42867T
Product # Size Price
42867T
1 Kit  (9 x 20 µl) $ 609

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 9664 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 17, 19 Rabbit IgG
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb 5625 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H Mk 89 Rabbit IgG
Phospho-RIP (Ser166) (D1L3S) Rabbit mAb 65746 20 µl
  • WB
H 78-82 Rabbit IgG
Phospho-RIP3 (Ser227) (D6W2T) Rabbit mAb 93654 20 µl
  • WB
  • IF
H 46-62 Rabbit IgG
Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb 91689 20 µl
  • WB
H 54 Rabbit IgG
Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb 36425 20 µl
  • WB
  • IP
  • IHC
H 30 Rabbit IgG
Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb 83186 20 µl
  • WB
  • IP
  • IF
H 17 Rabbit IgG
LC3B (E5Q2K) Mouse mAb 83506 20 µl
  • WB
  • IP
  • IHC
  • IF
H M R 14, 16 Mouse IgG2b
SQSTM1/p62 (D5L7G) Mouse mAb 88588 20 µl
  • WB
  • IP
  • IHC
  • IF
H 62 Mouse IgG1
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product Description

The Human Reactive Cell Death and Autophagy Antibody Sampler Kit provides an economical means of detecting common readouts in apoptosis, necroptosis, pyroptosis, and autophagy. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Each antibody in the Human Reactive Cell Death and Autophagy Antibody Sampler Kit detects endogenous levels of its target protein. Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb detects endogenous levels of the large fragment (17/19 kDa) of activated caspase-3 resulting from cleavage adjacent to Asp175. This antibody does not recognize full-length caspase-3 or other cleaved caspases. Non-specific labeling may be observed by immunofluorescence in specific sub-types of healthy cells in fixed-frozen tissues (e.g., pancreatic alpha-cells). Cytoplasmic background may be observed in human and monkey samples. Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb detects endogenous levels of the large fragment (89 kDa) of human PARP1 protein produced by caspase cleavage. The antibody does not recognize full-length PARP1 or other PARP isoforms. Cleaved Gasdermin D (Asp275) (E7H9G) Rabbit mAb recognizes endogenous levels of the amino-terminal fragment of Gasdermin D only when cleaved at Asp275. Cleaved-IL-1β (Asp116) (D3A3Z) Rabbit mAb recognizes endogenous levels of mature IL-1β protein only when cleaved at Asp116. Phospho-RIP3 (Ser227) (D6W2T) Rabbit mAb detected a band at 30 kDa that appears to be a cleavage product of RIP3. Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb may also bind to MLKL when dually phosphorylated at Thr357 and Ser358. LC3B (E5Q2K) Mouse mAb detects both type I and type II forms of LC3B. Cross reactivity was not detected with other family members.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp175 of human caspase-3, Asp214 of human PARP, Asp275 of human Gasdermin D, Asp116 of human IL-1β, Pro220 of human SQSTM1/p62, residues near the amino terminus of human LC3B, and synthetic phosphopeptides corresponding to Ser166 of human RIP, Ser227 of human RIP3, and Ser358 of human MLKL.

Background

Regulated cell death has been classified based on distinct morphological and biochemical pathways (1). Type I cell death, or apoptosis, is characterized by cytoplasmic shrinkage, chromatin condensation, nuclear fragmentation, plasma membrane blebbing, and phagocytic update of dead cells. Apoptosis can occur through extrinsic pathways involving extracellular factors, including the activation of death receptors, or through intrinsic pathways involving intracellular perturbations, including mitochondrial outer membrane permeabilization (2). Both of these apoptotic pathways lead to activation of caspases, a family of cysteine acid proteases that are synthesized as inactive zymogens containing pro-domains, followed by large (p20) and small (p10) subunits which are proteolytically activated in a cascade-like fashion. Caspase-3 is a key downstream protease activated by both extrinsic and intrinsic apoptotic pathways and cleaves a large number of proteins involved in the disassembly of the cell, including poly(ADP-ribose) polymerase (PARP), a protein involved in the DNA damage response.

 

Type II cell death, or autophagy, manifests with extensive cytoplasmic vacuolization, and like apoptosis, can include phagocytic update. Autophagy is a catabolic process for the degradation of cellular components including protein aggregates, damaged organelles, and pathogens (3). The process involves the engulfment of these components into a double membrane structure, the autophagosome, which fuses to the lysosome for degradation. Autophagy requires, and can be monitored by, the conversion of LC3 family members, such as LC3B, from a type I form to a lipidated type II form that is incorporated into the autophagosome membrane and binds to a variety of cargo receptors. Cargo receptors such as SQSTM1/p62 bind LC3 along with ubiquitinated proteins that are targeted for degradation. SQSTM1/p62 is also degraded during this process, and thus its expression is frequently used to monitor this process.

Type III cell death, or necrosis, manifests with plasma membrane permeability with cellular swelling and fragmentation, and lacks a clear phagocytic response which then leads to an inflammatory signaling with the release of damage-associated molecular patterns (DAMPs). Necrosis can be triggered by multiple regulated pathways including necroptosis and pyroptosis. Necroptosis is regulated by the kinase activities of RIP and RIP3 and the pore forming ability of MLKL (4). Necroptosis requires the activation of RIP3 which then phosphorylates MLKL at Ser358 (Ser345 in mouse). Phosphorylation of MLKL leads to generation of a pore complex involved in cell swelling and the secretion of DAMPs. RIP3 activation is triggered through several RIP homotypic interaction motif (RHIM) domain interactions including RIP, TRIF, and ZBP1 and results in the phosphorylation of RIP3 at Ser227 (Thr231/Ser232 in mouse). Canonical necroptosis signaling is mediated by RIP, and this can be inhibited by necrostatins, small molecules that directly inhibit RIP kinase activity. Activation of RIP can be monitored through autophosphorylation sites including Ser166. Pyroptosis is generally induced in cells of the innate immune system, and is characterized by cleavage of Gasdermin D (5). The amino-terminal fragment of Gasdermin D produced following cleavage by inflammatory caspases (Caspase-1, -4, -5), oligomerizes to form a pore. Canonical cleavage of Gasdermin D occurs through a two-step process. The first step involves transcriptional regulation of targets such as NLRP3 and the pro-forms of IL-1β and IL-18. In the second execution step, Caspase-1 is activated through formation of inflammasome complexes. Activated Caspase-1 cleaves Gasdermin D as well as IL-1β and IL-18 to their mature forms, and these active cytokines are secreted through pores formed by Gasdermin D.

  1. Galluzzi, L. et al. (2018) Cell Death Differ 25, 486-541.
  2. Green, D.R. (1998) Cell 94, 695-8.
  3. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ. 12 Suppl 2, 1509-18.
  4. Shan, B. et al. (2018) Genes Dev 32, 327-40.
  5. Shi, J. et al. (2017) Trends Biochem Sci 42, 245-54.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.