Product: Cleaved-Caspase 1 (Asp296), p20 Antibody
Catalog: AF4005
Source: Rabbit
Application: WB, IHC
Reactivity: Human, Mouse, Rat
Mol.Wt.: 20kD; 45kD(Calculated).
Uniprot: P29466
RRID: AB_2845463

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Clonality:
Polyclonal
Specificity:
Cleaved-Caspase-1 (Asp296,p20) Antibody detects endogenous levels of fragment of activated Caspase-1 resulting from cleavage adjacent to Asp296.
RRID:
AB_2845463
Cite Format: Affinity Biosciences Cat# AF4005, RRID:AB_2845463.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

CASP-1; CASP1; CASP1_HUMAN; Caspase 1; Caspase-1 subunit p10; ICE; IL-1 beta-converting enzyme; IL-1BC; IL1 beta converting enzyme; IL1B convertase; Interleukin 1 beta convertase; Interleukin 1B converting enzyme; Interleukin-1 beta convertase; Interleukin-1 beta-converting enzyme; p45;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P29466 CASP1_HUMAN:

Expressed in larger amounts in spleen and lung. Detected in liver, heart, small intestine, colon, thymus, prostate, skeletal muscle, peripheral blood leukocytes, kidney and testis. No expression in the brain.

Sequence:
MADKVLKEKRKLFIRSMGEGTINGLLDELLQTRVLNKEEMEKVKRENATVMDKTRALIDSVIPKGAQACQICITYICEEDSYLAGTLGLSADQTSGNYLNMQDSQGVLSSFPAPQAVQDNPAMPTSSGSEGNVKLCSLEEAQRIWKQKSAEIYPIMDKSSRTRLALIICNEEFDSIPRRTGAEVDITGMTMLLQNLGYSVDVKKNLTASDMTTELEAFAHRPEHKTSDSTFLVFMSHGIREGICGKKHSEQVPDILQLNAIFNMLNTKNCPSLKDKPKVIIIQACRGDSPGVVWFKDSVGVSGNLSLPTTEEFEDDAIKKAHIEKDFIAFCSSTPDNVSWRHPTMGSVFIGRLIEHMQEYACSCDVEEIFRKVRFSFEQPDGRAQMPTTERVTLTRCFYLFPGH

PTMs - P29466 As Substrate

Site PTM Type Enzyme
S16 Phosphorylation
T21 Phosphorylation
T32 Phosphorylation
K37 Ubiquitination
K44 Ubiquitination
T49 Phosphorylation
K53 Ubiquitination
K134 Ubiquitination
K148 Ubiquitination
S149 Phosphorylation
K158 Ubiquitination
K204 Ubiquitination
T226 Phosphorylation
S227 Phosphorylation
K268 Ubiquitination
K274 Ubiquitination
K278 Ubiquitination
S306 Phosphorylation
K319 Ubiquitination
K320 Ubiquitination
K325 Ubiquitination
S376 Phosphorylation

Research Backgrounds

Function:

Thiol protease that cleaves IL-1 beta between an Asp and an Ala, releasing the mature cytokine which is involved in a variety of inflammatory processes. Important for defense against pathogens. Cleaves and activates sterol regulatory element binding proteins (SREBPs). Can also promote apoptosis. Upon inflammasome activation, during DNA virus infection but not RNA virus challenge, controls antiviral immunity through the cleavage of CGAS, rendering it inactive. In apoptotic cells, cleaves SPHK2 which is released from cells and remains enzymatically active extracellularly.

PTMs:

The two subunits are derived from the precursor sequence by an autocatalytic mechanism.

Subcellular Location:

Cytoplasm. Cell membrane.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed in larger amounts in spleen and lung. Detected in liver, heart, small intestine, colon, thymus, prostate, skeletal muscle, peripheral blood leukocytes, kidney and testis. No expression in the brain.

Subunit Structure:

Heterotetramer that consists of two anti-parallel arranged heterodimers, each one formed by a 20 kDa (p20) and a 10 kDa (p10) subunit. The p20 subunit can also form a heterodimer with the epsilon isoform which then has an inhibitory effect. May be a component of the inflammasome, a protein complex which also includes PYCARD, CARD8 and NALP2 and whose function would be the activation of proinflammatory caspases. Both the p10 and p20 subunits interact with MEFV. Interacts with CARD17/INCA and CARD18. Interacts with SERPINB1; this interaction regulates CASP1 activity.

Family&Domains:

Belongs to the peptidase C14A family.

Research Fields

· Cellular Processes > Cell growth and death > Necroptosis.   (View pathway)

· Human Diseases > Neurodegenerative diseases > Amyotrophic lateral sclerosis (ALS).

· Human Diseases > Infectious diseases: Bacterial > Salmonella infection.

· Human Diseases > Infectious diseases: Bacterial > Pertussis.

· Human Diseases > Infectious diseases: Bacterial > Legionellosis.

· Human Diseases > Infectious diseases: Viral > Influenza A.

· Organismal Systems > Immune system > NOD-like receptor signaling pathway.   (View pathway)

· Organismal Systems > Immune system > Cytosolic DNA-sensing pathway.   (View pathway)

References

1). Jia X et al. Arsenic induces hepatic insulin resistance via mtROS-NLRP3 inflammasome pathway. J Hazard Mater 2020 Jun 4;399:123034. (PubMed: 32544768) [IF=14.224]

Application: WB    Species: Human    Sample: HepG2 cells

Fig.3 NaAsO2 impairs insulin signaling via activation of NLRP3 inflammasome in HepG2 cells. HepG2 cells were pretreated with 1 μg/ml LPS for 4 hours, 5 μM MCC950, for 4 hours, and were then treated with 4 μM NaAsO2 for 24 hours. Cytosolic fractions were analyzed by Western blot analysis. GAPDH was used as an internal control. The efficiency of MCC950, and its effect on NLRP3, caspase-1 activation, IL-1β and IL- 18 production in NaAsO2-treated HepG2 cells (A-E). HepG2 cells were stimulated with 100 nM insulin for 10 min at the end of treatment. Cytosolic fractions were analyzed by Western blot analysis. GAPDH was used as an internal control. The efficiency of MCC950, and its effect on p-IRS (Ser 307)/IRS1 and p-AKT (Ser473)/AKT1 ratio in NaAsO2-treated HepG2 cells (F-H). Insulin‐ stimulated glucose uptake in HepG2 cells was measured using a glucose assay kit (I). Results are mean ± SEM (n = 3). *P < 0.05 compare with the LPS group. #P<0.05 compare with 4 μM NaAsO2 group.

2). Chen Z et al. ELABELA attenuates deoxycorticosterone acetate/salt-induced hypertension and renal injury by inhibition of NADPH oxidase/ROS/NLRP3 inflammasome pathway. Cell Death Dis 2020 Aug 22;11(8):698. (PubMed: 32829380) [IF=9.685]

Application: IF/ICC    Species: mouse    Sample: HK2 cells

Fig. 7 Effects of ELA on the ROS production and NLRP3 imflammasome activation are independent of APJ. APJ knockdown was conducted by the transfection of plasmid pWHAPJg into HK2 cells and confirmed by RT-PCR. Then the HK2 cells with APJ knockdown were pretreated with ELA peptide (1 nM) for 1 h and then treated by Aldosterone (0.1 µM) for 24 h. a Representative immunoblots of APJ in renal cortex and medulla and summarized intensities of blots. b mRNA levels of APJ in HK2 cells with or without APJ knockdown. c ROS production in HK2 cells with APJ knockdown. d, e Representative confocal fluorescent images of co-localization (yellow) between Nlrp3 (green)/Caspase-1 (red) and summarized colocalization coefficient in HK2 cells with APJ knockdown. Scale bar, 10 µm. N = 6 per group. *P < 0.05 versus Ctrl group; #P < 0.05 versus DOCA/ salt group

3). Zhang Y et al. A unique death pathway keeps RIPK1 D325A mutant mice in check at embryonic day 10.5. PLoS Biol 2021 Aug 26;19(8):e3001304. (PubMed: 34437534) [IF=9.593]

4). Tao Y et al. Autophagic-CTSB-inflammasome axis modulates hepatic stellate cells activation in arsenic-induced liver fibrosis. Chemosphere 2019 Sep 24;242:124959 (PubMed: 31669990) [IF=8.943]

Application: WB    Species: rat    Sample: HSC-t6 cells

Fig.4 |NaAsO2-induced HSCs activation was dependent on the activation of NLRP3 inflammasome. (A, B) HSC-t6 cells were pretreated with or without MCC950 for 2h and then treated with NaAsO2 for 24 h. The expression level of caspase-1 p20, IL-1β, Collagen-, a-SMA, MMP-1, and TIMP-1 were determined by Western blot.

Application: WB    Species: rat    Sample: livers

Fig.2 NaAsO2 induced autophagy, activation of NLRP3 inflammasome and 662 hepatic stellate cell in vivo. (A) The expression level of LC3 and p62 in rat livers 663 following exposure to NaAsO2. The protein fraction was analyzed by Western blot. (B) 664 The expression level of cytoplasmic CTSB in rat livers following exposure to NaAsO2. 665 The protein fraction was analyzed by Western blot. (C) The expression level of NLRP3 inflammasome-associated proteins in rat livers following exposure to NaAsO2. 667 The protein fraction was analyzed by Western blot. (D) The expression level of 668 Collagen- , α-SMA, MMP-1, and TIMP-1in rat livers following exposure to NaAsO2. 669 The protein fraction was analyzed by Western blot. (E) α-SMA was detected by IF. 670 White arrows indicate positive staining. Values are mean ± SD, and n = 5. *p < 0.05, 671 **p < 0.01 compared with the control group; Scale bar = 100 µm

5). Ding R et al. Activating cGAS–STING axis contributes to neuroinflammation in CVST mouse model and induces inflammasome activation and microglia pyroptosis. J Neuroinflammation 2022 Jun 10;19(1):137. (PubMed: 35689216) [IF=8.322]

6). Du Y et al. The Dietary Supplement γ-Oryzanol Attenuates Hepatic Ischemia Reperfusion Injury via Inhibiting Endoplasmic Reticulum Stress and HMGB1/NLRP3 Inflammasome. Oxid Med Cell Longev 2021 Sep 2;2021:4628050. (PubMed: 34512864) [IF=7.310]

Application: WB    Species: Mice    Sample: liver tissue

Figure 5 ORY attenuated HMGB1/NLRP3 inflammasome during HIRI. (a, b) Proteins of hepatic tissues were determined by western blotting for the determination of HMGB1, NLRP3, cleaved-caspase-1, IL-1β, and GAPDH expressions. Relative protein abundance was semiquantified by densitometry (n = 4). Data were expressed as mean ± standard deviation (SD) values. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001 versus the sham group; #P < 0.05, ##P < 0.01, and ###P < 0.001 versus the I/R group; NS: no significance.

7). Zhang Q et al. Protective Effect of Yi Shen Pai Du Formula against Diabetic Kidney Injury via Inhibition of Oxidative Stress, Inflammation, and Epithelial-to-Mesenchymal Transition in db/db Mice. Oxid Med Cell Longev 2021 Aug 31;2021:7958021. (PubMed: 34504642) [IF=7.310]

Application: WB    Species: Mice    Sample: kidneys

Figure 6 YSPDF treatment regulated the expressions of TGF-β1, p-Smad2, Smad2, p-Smad3, Smad3, α-SMA, E-cadherin, Nrf2, HO-1, NQO1, NLRP3, ASC, caspase-1, and cleaved caspase-1 in the kidneys of db/db mice. Protein expression levels were normalized to the levels of either GAPDH or β-actin. The data were analyzed using a one-way ANOVA and expressed as the mean ± SEM (n = 3). #p < 0.05, ##p < 0.01, and ###p < 0.001 compared with the normal control group; ∗p < 0.05 and ∗∗p < 0.01 compared with the DKD group.

8). Du L et al. Novel biphenyl diester derivative AB-38b inhibits NLRP3 inflammasome through Nrf2 activation in diabetic nephropathy. Cell Biol Toxicol 2019 Nov 25 (PubMed: 31768838) [IF=6.819]

9). Sun Z et al. Inhibition of SGLT1 protects against glycemic variability-induced cardiac damage and pyroptosis of cardiomyocytes in diabetic mice. Life Sci 2021 Jan 25;119116. (PubMed: 33508297) [IF=6.780]

Application: WB    Species: mice    Sample: left ventricle tissues

Fig. 6. Knockdown of SGLT1 suppresses inflammatory response and pyroptosis in GVDM mice. A, Protein levels of NLRP3, ASC and caspase-1 (p20) in left ventricle tissues were measured by western blot. B, Quantitative analysis of relative protein levels of NLRP3, ASC and caspase-1. C, Protein levels of full length GSDMD and cleaved GSDMD were measured by western blot. D, Quantitative analysis of relative protein levels of cleaved GSDMD. E, The expression of NLRP3 was detected by immunofluorescence. Scale bar: 50 μm. F, Caspase-1 activity was measured by commercial kit. The relative mRNA levels of IL-1β (G) and IL-18 (H) were detected by qRT-PCR. I, The protein levels of IL-1β, pro-IL-1β and IL-18 were measured by western blot. Quantitative analysis of relative protein levels of IL-1β (J) and IL-18 (K). The release of IL-1β (L) and IL-18 (M) were detected by ELISA. Data were expressed as mean ± Standard deviation (SD). ***p < 0.001, **p < 0.01, *p < 0.05 vs control mice; †††p < 0.001, ††p < 0.01, † p < 0.05 vs DM mice; ###p < 0.001, ##p < 0.01 vs GVDM+shNC mice. DM, diabetes mellitus; GV, glycemic variability; SGLT1, sodium-glucose cotransporter 1; shSGLT1, SGLT1 shRNA; shNC, negative control shRNA; NLRP3, NLR family pyrin domain containing 3; ASC, apoptosis-associated speck-like protein containing CARD domain; GSDMD, gasdermin D.

10). Hong J et al. Bromodomain-containing protein 4 inhibition alleviates matrix degradation by enhancing autophagy and suppressing NLRP3 inflammasome activity in NP cells. J Cell Physiol 2020 Jan 24 (PubMed: 31975410) [IF=6.513]

Application: WB    Species: Human    Sample: NP cells

FIGURE 4 BRD4 inhibition alleviated NLRP3 inflammasome activity in NP cells. (a and b) Western blot (a) and subsequent densitometric analyses (b) revealed that NLRP3 inflammasome activity was significantly inhibited by JQ1. (c) Real‐time PCR was used to detect the effects of JQ1 on the induction of NLRP3 inflammasome–related gene expression by TNF‐α. (d and e) Western blot (d) and subsequent densitometric analyses (e) revealed the effect of JQ1 on the induction of NLRP3 inflammasome–related protein expression by TNF‐α. (f) Western blot demonstrated that NLRP3 inflammasome activity induction by TNF‐α was significantly alleviated by BRD4 knockdown with a lentivirus expressing shBRD4. The data represent the mean ± SD of three independent experiments. *p < .05. NP, nucleus pulposus; PCR, polymerase chain reaction; SD, standard deviation; TNF‐α, tumor necrosis factor α

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