The expression and purification of LpxA of Chlamydia trachomatis and preparation of its polyclonalantibody.
The aim of this research is to purify the LpxA protein of Chlamydia trachomatis (Ct) and put together the polyclonal antibody towards LpxA protein, in order to put a basis for learning the perform of LpxA protein. The LpxA gene was amplified by PCR. The expression plasmid pET28a-LpxA was constructed by utilizing pET28a because the vector.
The fusion protein containing 6 histidine tag was induced by IPTG and purified by Ni2+ chromatography gel. The purified His-LpxA protein was used as an immunogen to immunize New Zealand rabbits subcutaneously via the again to arrange polyclonal antibody. Immunoblotting was used to detect the response between the antibody and His-LpxA.
The willpower of polyclonal antibody titer was detected by ELISA. The relative molecular weight of His-LpxA was 32.eight kDa, and it may very well be expressed in Escherichia coli. The purity of the purified protein was about 95%. After immunizing New Zealand rabbits, the antiserum was in a position to acknowledge the recombinant His-LpxA protein with a titer larger than 1:10240. On this research, LpxA protein was efficiently purified and antiserum was ready, which supplied an experimental foundation for learning the perform of LpxA protein.
Description: This gene encodes a mitochondrial membrane protein that participates in mitochondrial fusion and contributes to the maintenance and operation of the mitochondrial network. This protein is involved in the regulation of vascular smooth muscle cell proliferation, and it may play a role in the pathophysiology of obesity. Mutations in this gene cause Charcot-Marie-Tooth disease type 2A2, and hereditary motor and sensory neuropathy VI, which are both disorders of the peripheral nervous system. Defects in this gene have also been associated with early-onset stroke. Two transcript variants encoding the same protein have been identified.
Description: Mfn2 is a protein encoded by the MFN2 gene which is approximately 86,4 kDa. Mfn2 is localised to the mitochondrion outer membrane. It is involved in pink/parkin mediated mitophagy, toll-like receptor signalling pathways and glucose / energy metabolism. It is a mitochondrial membrane protein that participates in mitochondrial fusion and contributes to the maintenance and operation of the mitochondrial network. Mfn2 is ubiquitously expressed at low levels. Mutations in the MFN2 gene may result in Charcot-Marie-Tooth disease. STJ94105 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen. This polyclonal antibody detects endogenous levels of Mfn2 protein.
Description: A polyclonal antibody against MFN2. Recognizes MFN2 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: WB, IHC, ELISA;WB:1/500-1/2000.IHC:1/100-1/300.ELISA:1/40000
Description: A polyclonal antibody against MFN2. Recognizes MFN2 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC
Description: A polyclonal antibody against MFN2. Recognizes MFN2 from Human, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC; Recommended dilution: WB:1:200-1:3000, IHC:1:20-1:200
Description: A polyclonal antibody for detection of Mfn2 from Human, Mouse, Rat. This Mfn2 antibody is for WB, IHC-P, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the Internal region of human Mfn2
Description: A polyclonal antibody for detection of Mfn2 from Human, Mouse, Rat. This Mfn2 antibody is for WB, IHC-P, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the Internal region of human Mfn2
Description: A polyclonal antibody for detection of Mfn2 from Human, Mouse, Rat. This Mfn2 antibody is for WB, IHC-P, ELISA. It is affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen synthesized peptide derived from the Internal region of human Mfn2
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human MFN2 . This antibody is tested and proven to work in the following applications:
Description: A polyclonal antibody against MFN2. Recognizes MFN2 from Human. This antibody is HRP conjugated. Tested in the following application: ELISA
Description: A polyclonal antibody against MFN2. Recognizes MFN2 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA
Description: A polyclonal antibody against MFN2. Recognizes MFN2 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human MFN2 (Center). This antibody is tested and proven to work in the following applications:
Description: Description of target: Essential transmembrane GTPase, which mediates mitochondrial fusion. Fusion of mitochondria occurs in many cell types and constitutes an important step in mitochondria morphology, which is balanced between fusion and fission. MFN2 acts independently of the cytoskeleton. It therefore plays a central role in mitochondrial metabolism and may be associated with obesity and/or apoptosis processes. Overexpression induces the formation of mitochondrial networks. Plays an important role in the regulation of vascular smooth muscle cell proliferation. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). Is required for PARK2 recruitment to dysfunctional mitochondria. Involved in the control of unfolded protein response (UPR) upon ER stress including activation of apoptosis and autophagy during ER stress. Acts as an upstream regulator of EIF2AK3 and suppresses EIF2AK3 activation under basal conditions.5 Publications
<p>Manually curated information for which there is published experimental evidence.</p>
<p><a href="/manual/evidences#ECO:0000269">More…</a></p> Manual assertion based on experiment iniRef.2"Dysregulation of HSG triggers vascular proliferative disorders."_x005F_x005F_x000D_Chen K.-H., Guo X., Ma D., Guo Y., Li Q., Yang D., Li P., Qiu X., Wen S., Xiao R.-P., Tang J._x005F_x005F_x000D_Nat. Cell Biol. 6:872-883(2004) [PubMed] [Europe PMC] [Abstract]Cited for: NUCLEOTIDE SEQUENCE [MRNA] (ISOFORM 1), FUNCTION, DISEASE.Ref.9"Control of mitochondrial morphology by a human mitofusin."_x005F_x005F_x000D_Santel A., Fuller M.T._x005F_x005F_x000D_J. Cell Sci. 114:867-874(2001) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION, SUBCELLULAR LOCATION, MUTAGENESIS OF LYS-109; 622-GLY--VAL-624 AND 657-LYS--ARG-659.Ref.10"Membrane topology and mitochondrial targeting of mitofusins, ubiquitous mammalian homologs of the transmembrane GTPase Fzo."_x005F_x005F_x000D_Rojo M., Legros F., Chateau D., Lombes A._x005F_x005F_x000D_J. Cell Sci. 115:1663-1674(2002) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION, SUBCELLULAR LOCATION, MEMBRANE TOPOLOGY, TISSUE SPECIFICITY, MUTAGENESIS OF LYS-109; SER-110 AND ARG-259.Ref.16"MiD49 and MiD51 can act independently of Mff and Fis1 in Drp1 recruitment and are specific for mitochondrial fission."_x005F_x005F_x000D_Palmer C.S., Elgass K.D., Parton R.G., Osellame L.D., Stojanovski D., Ryan M.T._x005F_x005F_x000D_J. Biol. Chem. 288:27584-27593(2013) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION.Ref.17"PINK1-phosphorylated mitofusin 2 is a Parkin receptor for culling damaged mitochondria."_x005F_x005F_x000D_Chen Y., Dorn G.W. II_x005F_x005F_x000D_Science 340:471-475(2013) [PubMed] [Europe PMC] [Abstract]Cited for: FUNCTION IN MITOPHAGY, INTERACTION WITH PARK2, PHOSPHORYLATION AT THR-111 AND SER-442 BY PINK1, UBIQUITINATION BY PARK2, SUBCELLULAR LOCATION, MUTAGENESIS OF THR-111 AND SER-442. ;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich Immunoassay;Sensitivity: < 0.055 ng/mL
Description: Description of target: This gene encodes a mitochondrial membrane protein that participates in mitochondrial fusion and contributes to the maintenance and operation of the mitochondrial network. This protein is involved in the regulation of vascular smooth muscle cell proliferation, and it may play a role in the pathophysiology of obesity. Mutations in this gene cause Charcot-Marie-Tooth disease type 2A2, and hereditary motor and sensory neuropathy VI, which are both disorders of the peripheral nervous system. Defects in this gene have also been associated with early-onset stroke. Two transcript variants encoding the same protein have been identified.;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 35pg/mL
Efficiency analysis of the polyclonal anti-rabies virus ribonucleoprotein IgG antibodies produced in-house to be used in direct fluorescent antibody check.
Fluorescein isothiocyanate (FITC) labelled anti-rabies virus ribonucleoprotein (RNP) antibodies can be utilized as immunoreagents in direct fluorescent antibody testing (dFAT) for rabies diagnoses. Whereas in-house merchandise are sometimes utilized by laboratories, most conjugates are business reagents. Business anti-RNP antibodies are solely accessible for analysis functions in Brazil, nevertheless, which contributes to the rising use of in-house produced antibodies.
Contemplating that conjugate high quality might affect the outcomes obtained throughout rabies analysis, we sought to investigate the efficiency necessities of in-house produced polyclonal anti-RNP IgG-FITC for utility in dFAT. To that finish, their reproducibility, diagnostic sensitivity, and specificity have been evaluated. The titer of polyclonal anti-RNP IgG-FITC was initially decided and evaluated by dFAT, utilizing central nervous system (CNS) samples of various animal species (canines, cats, bovines, equines, bats, and non-human primates).
As our predominant end result, the polyclonal anti-RNP IgG-FITC reached a titer of 1:30/1:40 in dFAT, with 100% of diagnostic sensitivity and specificity. When it comes to reproducibility, the antibodies, regardless the manufacturing batch, introduced the identical performances. In conclusion, the in-house produced polyclonal anti-RNP IgG-FITC proved appropriate for rabies virus antigen detection by dFAT.
Manufacturing of F(ab’)2 from Monoclonal and PolyclonalAntibodies.
Antibodies are broadly utilized in therapeutic, diagnostic, and analysis purposes, and antibody derivatives reminiscent of F(ab’)2 fragments are used when solely a specific antibody area is required. F(ab’)2 will be produced via antibody engineering, however some purposes require F(ab’)2 produced from an authentic formulated antibody or instantly from a polyclonal antibody pool.
The cysteine protease immunoglobulin-degrading enzyme (IdeS) from Streptococcus pyogenes digests immunoglobulin G (IgG) particularly and effectively to supply F(ab’)2 . Right here we element the manufacturing and purification of recombinant IdeS; its utilization to digest monoclonal or polyclonal antibodies to F(ab’)2 fragments; and F(ab’)2 purification via consecutive affinity chromatography steps.
Analysis of liquid and powdered types of polyclonalantibody preparation towards Streptococcus bovis and Fusobacterium necrophorum in cattle tailored or not tailored to extremely fermentable carbohydrate diets.
Feed components that modify rumen fermentation can be utilized to forestall metabolic disturbances reminiscent of acidosis and optimize beef cattle manufacturing. The research evaluated the consequences of liquid and powdered types of polyclonal antibody preparation (PAP) towards Streptococcus bovis and Fusobacterium necrophorum on rumen fermentation parameters in ruminally cannulated non-lactating dairy cows that have been tailored or unadapted to a excessive focus weight-reduction plan.
A double 3 × Three Latin sq. design was used with three PAP therapies (management, powdered and liquid PAP) and two adaptation protocols (tailored, unadapted; utilized to the sq.). Tailored animals have been transitioned for two weeks from an all-forage to an 80% focus weight-reduction plan, whereas unadapted animals have been switched abruptly.
Interactions between sampling time and adaptation have been noticed; 12 h after feeding, the tailored group had decrease ruminal pH and larger complete brief chain fatty acid concentrations than the unadapted group, whereas the alternative was noticed after 24 h. Acetate:propionate ratio, molar proportion of butyrate and ammonia nitrogen focus have been typically larger in tailored than unadapted cattle as much as 36 h after feeding.
Adaptation promoted 3.5 instances the variety of Entodinium protozoa however copy numbers of S. bovis and Fibrobacter succinogens genes in rumen fluid weren’t affected. Nonetheless, neither liquid nor powdered types of PAP altered rumen acidosis variables in tailored or unadapted animals.Adaptation of cattle to extremely fermentable carbohydrate diets promoted a extra secure ruminal setting, however PAP was not efficient on this research by which no animal skilled acute or sub-acute rumen acidosis.
Description: CpG methylation is an epigenetic modification that is important for embryonic development, imprinting, and X-chromosome inactivation. Studies in mice have demonstrated that DNA methylation is required for mammalian development. This gene encodes a DNA methyltransferase that is thought to function in de novo methylation, rather than maintenance methylation. The protein localizes to the cytoplasm and nucleus and its expression is developmentally regulated.
Description: CpG methylation is an epigenetic modification that is important for embryonic development, imprinting, and X-chromosome inactivation. Studies in mice have demonstrated that DNA methylation is required for mammalian development. This gene encodes a DNA methyltransferase that is thought to function in de novo methylation, rather than maintenance methylation. The protein localizes to the cytoplasm and nucleus and its expression is developmentally regulated.
Description: A sandwich quantitative ELISA assay kit for detection of Human DNA Methyltransferase 3A (DNMT3A) in samples from tissue homogenates, cell lysates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Human DNA Methyltransferase 3A (DNMT3A) in samples from tissue homogenates, cell lysates or other biological fluids.
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC;ELISA:1:2000-1:10000, WB:1:1000-1:5000, IHC:1:100-1:300
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC;ELISA:1:2000-1:5000, IHC:1:50-1:200
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC;ELISA:1:2000-1:5000, IHC:1:100-1:300
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC;ELISA:1:2000-1:5000, IHC:1:50-1:200
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, IHC
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human, Mouse. This antibody is Unconjugated. Tested in the following application: ELISA, IHC, IF; Recommended dilution: IHC:1:20-1:200, IF:1:50-1:200
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human. This antibody is HRP conjugated. Tested in the following application: ELISA
Description: A polyclonal antibody against DNMT3A. Recognizes DNMT3A from Human. This antibody is FITC conjugated. Tested in the following application: ELISA
Soluble Epoxide Hydrolase Detection by Combining a Polyclonal Capture Antibody
Improvement of a Extremely Delicate Enzyme-Linked Immunosorbent Assay for Mouse Soluble Epoxide Hydrolase Detection by Combining a Polyclonal Seize Antibody with a Nanobody Tracer...
Utero Exposure To Endogenous Maternal Polyclonal Anti-Caspr2 antibody leads
In utero publicity to endogenous maternal polyclonal anti-Caspr2 antibody results in behavioral abnormalities resembling autism spectrum dysfunction in male mice The idea that...
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