Activation of toll like receptor 4 (TLR4) promotes cardiomyocyte apoptosis through SIRT2 dependent p53 deacetylation
December 11, 2020

Activation of toll like receptor 4 (TLR4) promotes cardiomyocyte apoptosis through SIRT2 dependent p53 deacetylation

By Dylan

Cardiomyocyte irritation adopted by apoptosis and fibrosis is a crucial mediator for growth and development of coronary heart failure. Activation of toll-like receptor 4 (TLR4), an necessary regulator of irritation, causes the development of cardiac hypertrophy and harm. Nevertheless, the exact mechanism of TLR4-mediated antagonistic cardiac outcomes remains to be elusive. The current research was designed to seek out the function of TLR4 in cardiac fibrosis and apoptosis, and molecular mechanism thereof. Rats have been handled with TLR4 agonist (LPS 12.5 μg/kg/day) via osmotic pump for <em>14</em> days. To simulate the situation in vitro, H9c2 cells have been handled with LPS (1 μg/ml). Equally, H9c2 cells have been transfected with TLR4 and SIRT2 c-DNA clone for overexpression. Myocardial oxidative stress, irritation, fibrosis and mitochondrial parameters have been evaluated each in vitro and in vivo. Cardiac irritation after LPS therapy was confirmed by elevated TNF-α and IL-6 expression in rat coronary heart.

There was a marked improve in oxidative stress as noticed by elevated TBARS and decreased endogenous antioxidants (GSH and catalase), together with mitochondrial dysfunction as measured by mitochondrial complicated exercise in LPS-treated rat hearts. Histopathological examination confirmed the presence of cardiac fibrosis after LPS therapy. Protein expression of nuclear p53 and cleaved <em>caspase</em>-7/<em>caspase</em>-9 was considerably elevated in LPS handled coronary heart. Just like in vivo research, nuclear translocation of p53, mitochondrial dysfunction and mobile apoptosis have been noticed in H9c2 cells handled with LPS. Our knowledge additionally point out that decreased expression of SIRT2 was related to elevated acetylation of p53 after LPS therapy. In conclusion, TLR4 activation in rats promotes cardiac irritation, mitochondrial dysfunction, apoptosis and fibrosis. p53 and <em>caspase</em> 7/<em>caspase</em> 9 have been discovered to play an necessary function in TLR4-mediated apoptosis.

Our knowledge counsel that, lowering TLR4 mediated fibrosis and apoptosis may very well be a novel method within the therapy of coronary heart failure, retaining within the view the main function performed by TLR4 in cardiac irritation. BACKGROUND Cardiac distant ischemic conditioning (RIC) is a noninvasive cardioprotective methodology in ischemia-reperfusion harm and acute myocardial infarction (AMI). The goals of this research have been to analyze the results of RIC in a rat mannequin of AMI. MATERIAL AND METHODS Grownup male Sprague-Dawley rats included the AMI group that underwent ligation of the left anterior descending (LAD) coronary artery (n=24), the RIC group that consisted the AMI rat mannequin handled with RIC as soon as each day within the left hind limb till days 1, 7 and <em>14</em> (n=24), and the sham group (n=24). Myocardial infarct measurement was measured by routine histology with triphenyltetrazolium chloride (TTC) and Masson’s trichrome histochemical staining for myocardial necrosis and fibrosis, respectively.

 

Mitochondrial defect in muscle precedes neuromuscular junction degeneration and motor neuron dying in CHCHD10S59L/+ mouse.

Lately, we offered genetic foundation exhibiting that mitochondrial dysfunction can set off motor neuron degeneration, via identification of CHCHD10 encoding a mitochondrial protein. We reported sufferers, carrying the p.Ser59Leu heterozygous mutation in CHCHD10, from a big household with a mitochondrial myopathy related to motor neuron illness (MND). Quickly, our group and others reported CHCHD10 mutations in amyotrophic lateral sclerosis (ALS), frontotemporal dementia-ALS and different neurodegenerative illnesses. Right here, we generated knock-in (KI) mice, carrying the p.Ser59Leu mutation, that mimic the mitochondrial myopathy with mtDNA instability displayed by the sufferers from our authentic household. Earlier than months of age, all KI mice developed a deadly mitochondrial cardiomyopathy related to enhanced mitophagy.
mice additionally displayed neuromuscular junction (NMJ) and motor neuron degeneration with hyper-fragmentation of the motor finish plate and average however vital motor neuron loss in lumbar spinal twine on the finish stage of the illness. At this stage, we noticed TDP-43 cytoplasmic aggregates in spinal neurons. We additionally confirmed that motor neurons differentiated from human iPSC carrying the p.Ser59Leu mutation have been way more delicate to Staurosporine or glutamate-induced <em>caspase</em> activation than management cells. These knowledge verify that mitochondrial deficiency related to CHCHD10 mutations may be on the origin of MND. CHCHD10 is extremely expressed within the NMJ post-synaptic half. Importantly, the fragmentation of the motor finish plate was related to irregular CHCHD10 expression that was additionally noticed closed to NMJs which have been morphologically regular.
Moreover, we discovered OXPHOS deficiency in muscle of CHCHD10 mice at Three months of age within the absence of neuron loss in spinal twine. Our knowledge present that the pathological results of the p.Ser59Leu mutation goal muscle previous to NMJ and motor neurons. They seemingly result in OXPHOS deficiency, lack of cristae junctions and destabilization of inside membrane construction inside mitochondria at motor finish plate of NMJ, impairing neurotransmission. These knowledge are in favor with a key function for muscle in MND related to CHCHD10 mutations.
 Activation of toll like receptor 4 (TLR4) promotes cardiomyocyte apoptosis through SIRT2 dependent p53 deacetylation

Activation of toll like receptor 4 (TLR4) promotes cardiomyocyte apoptosis through SIRT2 dependent p53 deacetylation

Manganese suppresses oxidative stress, irritation and caspase-Three activation in rats uncovered to chlorpyrifos.

The current research investigated the person and mixed impression of organophosphorus pesticide chlorpyrifos (CPF) and manganese (Mn), a naturally occurring hint steel, on hepatorenal perform in grownup rats. The 4 experimental teams particularly management, CPF alone (5 mg/kg), Mn alone (10 mg/kg) and the co-exposure group consisted of eight rats which have been orally gavage for <em>14</em> consecutive days. Following sacrifice, the biomarkers of hepatorenal injury, antioxidant enzyme actions, myeloperoxidase (MPO) exercise in addition to ranges of nitric oxide, reactive oxygen and nitrogen (RONS) species and lipid peroxidation (LPO) have been analysed spectrophotometrically.
Additional, the focus of tumour necrosis issue alpha (TNF-α), interleukin-1 β (IL-1β) and <em>caspase</em>-Three exercise have been assessed utilizing ELISA. Outcomes confirmed that the CPF-induced improve in biomarkers of hepatorenal toxicity have been considerably (p < 0.05) alleviated in rats co-expose to CPF and Mn. Furthermore, the lower in antioxidant standing in addition to the elevation in RONS and LPO have been considerably assuaged in rats co-treated with CPF and Mn. As well as, CPF mediated improve in TNF-α, IL-1β and <em>caspase</em>-Three exercise have been considerably diminished within the liver and kidney of rats co-exposed to CPF and Mn.

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Description: Caspase-1 is a protein encoded by the CASP1 gene which is approximately 45,1 kDa. Caspase-1 is localised to the cytoplasm. It is involved in amyotrophic lateral sclerosis, toll-like receptor signalling pathways, p53 signalling and the TNFR1 pathway. This protein falls under the the cysteine-aspartic acid protease family. upon activation It plays a central role in the execution-phase of cell apoptosis. These enzymes can be divided into initiators and effectors. The initiator isoforms are activated by, and interact with, upstream adaptor molecules. Caspase-1 is expressed mainly in the spleen and lung. Mutations in the CASP1 gene may result in shigellosis and cowpox. STJ98574 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of Caspase-1.

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Description: Caspase-1 is a protein encoded by the CASP1 gene which is approximately 45,1 kDa. Caspase-1 is localised to the cytoplasm. It is involved in amyotrophic lateral sclerosis, toll-like receptor signalling pathways, p53 signalling and the TNFR1 pathway. This protein falls under the the cysteine-aspartic acid protease family. upon activation It plays a central role in the execution-phase of cell apoptosis. These enzymes can be divided into initiators and effectors. The initiator isoforms are activated by, and interact with, upstream adaptor molecules. Caspase-1 is expressed mainly in the spleen and lung. Mutations in the CASP1 gene may result in shigellosis and cowpox. STJ90001 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of fragment activated Caspase-1 protein resulting from cleavage adjacent to D210.

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Polyclonal Goat anti-GST μ-form

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Caspase-1 Antibody

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Caspase-1 Antibody

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anti- Caspase 6 antibody

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anti- Caspase 8 antibody

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anti- Caspase 8 antibody

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Description: Antibody raised against Caspase 9

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Description: Antibody raised against Caspase 9

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Anti-Caspase-4 antibody

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EUR 332.4

Anti-Caspase-9 antibody

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EUR 471.6
Description: This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein can undergo autoproteolytic processing and activation by the apoptosome, a protein complex of cytochrome c and the apoptotic peptidase activating factor 1; this step is thought to be one of the earliest in the caspase activation cascade. This protein is thought to play a central role in apoptosis and to be a tumor suppressor. Alternative splicing results in multiple transcript variants.

Anti-Caspase-3 antibody

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Description: This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein cleaves and activates caspases 6, 7 and 9, and the protein itself is processed by caspases 8, 9 and 10. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease. Alternative splicing of this gene results in two transcript variants that encode the same protein.

Anti-Caspase-3 antibody

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Description: This gene encodes a protein which is a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein cleaves and activates caspases 6, 7 and 9, and the protein itself is processed by caspases 8, 9 and 10. It is the predominant caspase involved in the cleavage of amyloid-beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease. Alternative splicing of this gene results in two transcript variants that encode the same protein.

Anti-Caspase-2 antibody

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Description: Rabbit polyclonal to Caspase-2.

Anti-Caspase-3 antibody

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EUR 236.4
Description: Rabbit polyclonal to Caspase-3.

Anti-Caspase-4 antibody

STJ92022 200 µl
EUR 236.4
Description: Caspase-4 is a protein encoded by the CASP4 gene which is approximately 43,2 kDa. Caspase-4 is localised to the cytoplasm, cytosol and endoplasmic reticulum membrane. It is involved in the p53 signalling, the TRAF pathway, ceramide pathway and innate immune system. CASP4 is an essential effector of NLRP3 inflammasome-dependent CASP1 activation and IL1B and IL18 secretion in response to non-canonical activators, such as UVB radiation. It is involved in the signalling pathways of apoptosis, necrosis and inflammation. It is widely expressed, including keratinocytes and colonic and small intestinal epithelial cells. Mutations in the CASP4 gene may result in neuroblastoma. STJ92022 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of Caspase-4 protein.

Anti-Caspase-6 antibody

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Description: Rabbit polyclonal to Caspase-6.

Anti-Caspase-7 antibody

STJ92024 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-7.

Anti-Caspase-8 antibody

STJ92025 200 µl
EUR 236.4
Description: Caspase-8 is a protein encoded by the CASP8 gene which is approximately 55,3 kDa. Caspase-8 is localised to the cytoplasm and is involved in the TNFR1 pathway, dimerization of procaspase-8 , activated TLR4 signalling, apoptosis signalling and toll-like receptor signalling pathways. This protein falls under the cysteine-aspartic acid protease family. It pays a role in the programmed cell death induced by Fas and various apoptotic stimuli. Caspase-8 isoform 1, 5 and 7 are expressed in a wide variety of tissues. Mutations in the CASP8 gene may result in a caspase-8 deficiency. STJ92025 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of Caspase-8 protein.

Anti-Caspase-9 antibody

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Description: Rabbit polyclonal to Caspase-9.

Anti-Caspase-9 antibody

STJ92027 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-9.

Anti-Caspase-9 antibody

STJ92028 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-9.

Anti-Caspase-9 antibody

STJ92029 200 µl
EUR 236.4
Description: Caspase-9 is a protein encoded by the CASP9 gene which is approximately 46,2 kDa. Caspase-9 is localised to the cytosol, mitochondrion and nucleus. It is involved in the TNFR1 pathway, RET signalling and apoptosis modulation and signalling. Caspase-9 exist as an inactive proenzyme which undergoes proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. It is involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Caspase-9 is ubiquitously expressed with highest expression in the heart. Mutations in the CASP9 gene may result in cerebral hypoxia and cervical cancer. STJ92029 was affinity purified. This poly clonal antibody binds endogenous caspase-9.

Anti-Caspase-3 antibody

STJ98967 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-3.

Anti-Caspase-9 antibody

STJ99219 200 µl
EUR 236.4
Description: Mouse monoclonal to Caspase-9.

Anti-Caspase 3 antibody

STJ99611 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase 3.

Anti-Caspase-10 antibody

STJ96459 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-10.

Anti-Caspase-3 antibody

STJ96569 200 µl
EUR 236.4
Description: Caspase-3 is a protein encoded by the CASP3 gene which is approximately 31,6 kDa. Caspase-3 is localised to the cytoplasm. It is involved in the TNFR1 pathway, apoptosis modulation and signalling and respiratory electron transport. This protein falls under the cysteine-aspartic acid protease family. It is a sequential activatior of caspases and plays a central role in the execution-phase of cell apoptosis. Caspase-3 is highly expressed in lung, spleen, heart, liver and kidney. Mutations in the CASP3 gene may result in bladder urothelial carcinoma and primary effusion lymphoma. STJ96569 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of fragment of Caspase-3 protein.

Anti-Caspase 9 antibody

STJ96979 200 µl
EUR 236.4
Description: Caspase-9 is a protein encoded by the CASP9 gene which is approximately 46,2 kDa. Caspase-9 is localised to the cytosol, mitochondrion and nucleus. It is involved in the TNFR1 pathway, RET signalling and apoptosis modulation and signalling. Caspase-9 exist as an inactive proenzyme which undergoes proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. It is involved in the activation cascade of caspases responsible for apoptosis execution. Binding of caspase-9 to Apaf-1 leads to activation of the protease which then cleaves and activates caspase-3. Caspase-9 is ubiquitously expressed with highest expression in the heart. Mutations in the CASP9 gene may result in cerebral hypoxia and cervical cancer. STJ96979 was affinity purified. This monoclonal antibody binds endogenous caspase-9.

Anti-Caspase-8 antibody

STJ97394 200 µl
EUR 236.4
Description: Caspase-8 is a protein encoded by the CASP8 gene which is approximately 55,3 kDa. Caspase-8 is localised to the cytoplasm and is involved in the TNFR1 pathway, dimerization of procaspase-8, activated TLR4 signalling, apoptosis signalling and toll-like receptor signalling pathways. This protein falls under the cysteine-aspartic acid protease family. It pays a role in the programmed cell death induced by Fas and various apoptotic stimuli. Caspase-8 isoform 1, 5 and 7 are expressed in a wide variety of tissues. Mutations in the CASP8 gene may result in a caspase-8 deficiency. STJ97394 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of Caspase-8 protein.

Anti-Caspase-3 antibody

STJ97452 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-3.

Anti-Caspase-8 antibody

STJ97456 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-8.

Anti-Caspase-3 antibody

STJ97723 200 µl
EUR 236.4
Description: Mouse monoclonal to Caspase-3.

Anti-Caspase-3 antibody

STJ97730 200 µl
EUR 236.4
Description: Mouse monoclonal to Caspase-3.

Anti-Caspase-8 antibody

STJ97895 100 µl
EUR 280.8
Description: Mouse monoclonal to Caspase-8.

Anti-Caspase-3 antibody

STJ119900 100 µl
EUR 471.6

Anti-caspase-4 antibody

STJ16101312 100 µg
EUR 826.8

Anti-caspase-7 antibody

STJ16101313 100 µg
EUR 826.8

Anti-caspase-4 antibody

STJ16101448 100 µg
EUR 826.8

Anti-caspase-7 antibody

STJ16101449 100 µg
EUR 826.8

Anti-Caspase 6 antibody

STJ73022 100 µg
EUR 430.8

Anti-Caspase-14 antibody

STJ90022 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-14.

Anti-Caspase-3 antibody

STJ90025 200 µl
EUR 236.4
Description: Caspase-3 is a protein encoded by the CASP3 gene which is approximately 31,6 kDa. Caspase-3 is localised to the cytoplasm. It is involved in the TNFR1 pathway, apoptosis modulation and signalling and respiratory electron transport. This protein falls under the cysteine-aspartic acid protease family. It is a sequential activatior of caspases and plays a central role in the execution-phase of cell apoptosis. Caspase-3 is highly expressed in lung, spleen, heart, liver and kidney. Mutations in the CASP3 gene may result in bladder urothelial carcinoma and primary effusion lymphoma. STJ90025 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of fragment of Caspase-3 protein.

Anti-Caspase-7 antibody

STJ90031 200 µl
EUR 236.4
Description: Caspase-7 is a protein encoded by the CASP7 gene which is approximately 34,2 kDa. Caspase-7 is localised to the cytoplasm. It is involved in the TNFR1 pathway and apoptosis modulation and signalling. It exists as an inactive proenzyme which undergoes proteolytic processing at conserved aspartic residues to produce two subunits that dimerize to form the active enzyme. It is involved in the activation cascade of caspases responsible for apoptosis execution. Caspase-7 is highly expressed in the lung, skeletal muscle, liver, kidney, spleen and heart. Mutations in the CASP7 gene may result in breast adenocarcinoma. STJ90031 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of Caspase-7 protein.

Anti-Caspase-6 antibody

STJ90104 200 µl
EUR 236.4
Description: Rabbit polyclonal to Caspase-6.

Anti-Caspase 3 antibody

STJ70944 100 µg
EUR 430.8

Anti-Caspase-3 Antibody

A1751-100 each
EUR 574.8

Anti-Caspase-7 Antibody

A1752-100 each
EUR 574.8

Histone H3 Methylation Antibody Panel Pack I – Active Genes

C10000
  • EUR 928.26
  • EUR 588.50
  • 4 x 25 µg
  • 4 x 25 ul

Histone H3 Methylation Antibody Panel Pack I – Repression Genes

C10001
  • EUR 928.26
  • EUR 588.50
  • 4 x 25 µg
  • 4 x 25 ul

Histone H3 Methylation Antibody Panel Pack II – Active Genes

C10002
  • EUR 754.26
  • EUR 470.80
  • 3 x 25 µg
  • 3 x 25 ul

Histone H3 Methylation Antibody Panel Pack II – Repression Genes

C10003
  • EUR 928.26
  • EUR 588.50
  • 4 x 25 µg
  • 4 x 25 ul

Histone H3 Methylation Antibody Panel Pack III – Active Genes

C10004
  • EUR 928.26
  • EUR 588.50
  • 4 x 25 µg
  • 4 x 25 ul

Caspase Inhibitor Set I

A9901-1 1 Set
EUR 184.8

Caspase-3/7 Inhibitor

2780-1 each
EUR 183.6

Anti-EDG-1 Antibody

A01502-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for EDG-1 Antibody (S1PR1) detection.tested for WB in Human, Mouse, Rat.

Anti-ROBO-1 Antibody

A01530-1 100ug
EUR 546
Description: Rabbit Polyclonal ROBO-1 Antibody. Validated in IF, IHC, WB and tested in Human, Mouse, Rat.

Anti-Bag-1 Antibody

A02423-1 50 ul
EUR 476.4
Description: Rabbit Polyclonal Bag-1 Antibody. Validated in IP, IHC and tested in Bovine, Canine, Human, Mouse, Rat.

Anti-TUB 1 Antibody

A02917-1 100ug/vial
EUR 352.8

Anti-Dok-1 Antibody

A03039-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for Dok-1 Antibody (DOK1) detection.tested for WB in Human, Mouse, Rat.

Anti-TFIIIB90-1 Antibody

A03761-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for TFIIIB90-1 Antibody (BRF1) detection.tested for WB in Human, Mouse.

Anti-Atrophin-1 Antibody

A03828-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for Atrophin-1 Antibody (ATN1) detection. Tested with WB in Human, Mouse, Rat.

Anti-CNG-1 Antibody

A05494-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for CNG-1 Antibody (CNGA1) detection. Tested with WB in Human, Mouse, Rat.

Anti-Periphilin 1 Antibody

A06996-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for Periphilin 1 Antibody (PPHLN1) detection.tested for WB in Human, Mouse.

Anti-Lyl-1 Antibody

A07491-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Lyl-1 Antibody. Validated in IHC and tested in Human, Mouse, Rat.

Anti-GLI-1 Antibody

A07972-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for GLI-1 Antibody (ACSS1) detection. Tested with WB in Human, Mouse, Rat.

Anti-Cerebellin 1 Antibody

A09176-1 100ul
EUR 476.4
Description: Rabbit Polyclonal Antibody for Cerebellin 1 Antibody (CBLN1) detection. Tested with WB in Human, Mouse, Rat.
Mild microscopic examination evidenced that the severity of histopathological lesions induced by CPF have been alleviated in rats co-exposed to CPF and Mn. In conclusion, the outcomes spotlight that co-exposure to CPF and Mn in rats assuaged CPF-induced oxidative stress, irritation and <em>caspase</em>-Three activation within the liver and kidney of the rats.