Current research of grownup neurogenesis of the mammalian central nervous system have instructed sudden plasticity and complexity of neural cell ontogenesis. Redefinition and reconstitution of cell classification and lineage relationships, particularly between glial and neural precursors, are an pressing and essential concern.
Within the current examine, we describe a brand new monoclonal antibody, A3B10, which was produced by immunizing mice with the membrane fraction ready from astrocyte-enriched major neural cell cultures. Immunohistochemistry of mind sections, together with brains from glial fibrillary acidic protein (GFAP)-deficient mice and first combined neural cell cultures, in addition to immunoblot evaluation and immunoelectron microscopy, have revealed that 1) A3B10 acknowledges a majority of cells in ependyma in neonatal and grownup rats, 2) A3B10 stains nearly all GFAP(+) cells and a few S100beta(+) cells within the corpus callosum, 3) A3B10 particularly stains astrocytes in vitro in major cultures of rat embryonic cerebral hemispheres, 4) A3B10 equally stains ependymal cells of wild-type and GFAP-deficient mice, and 5) A3B10 antigen would possibly assemble intermediate filament bundles with GFAP and/or vimentin.
These information instructed that the antibody labels a big selection of astorcytic-lineage cells together with astrocytes, astrocyte precursors, and neural stem cells. Screening a cDNA library derived from rat embryonic mind has revealed that the antibody acknowledges calmodulin-regulated spectrin-associated protein 1 (Camsap1). Thus this antibody could present not solely a brand new marker to establish astrocyte-lineage cells but in addition a brand new goal molecule to elucidate the ontogeny, improvement, and pathophysiological features of astrocyte-lineage cells.
Potent neutralization of anthrax edema toxin by a humanized monoclonal antibody that competes with calmodulin for edema issue binding.
This examine describes the isolation and characterization of a neutralizing monoclonal antibody (mAb) in opposition to anthrax edema issue, EF13D. EF13D neutralized edema toxin (ET)-mediated cyclic AMP (cAMP) responses in cells and guarded mice from each ET-induced footpad edema and systemic ET-mediated lethality. The antibody epitope was mapped to area IV of EF.
The mAb was in a position to compete with calmodulin (CaM) for EF binding and displaced CaM from EF-CaM complexes. EF-mAb binding affinity (0.05-0.12 nM) was 50- to 130-fold increased than that reported for EF-CaM. This anti-EF neutralizing mAb may probably be used alone or with an anti-PA mAb within the emergency prophylaxis and remedy of anthrax an infection.
Manufacturing of calmodulin-tagged proteins in Drosophila Schneider S2 cells: a novel system for antigen manufacturing and phage antibody isolation.
We report the event of an expression system for the manufacturing of soluble, calmodulin (CaM)-tagged proteins in Drosophila Schneider S2 cells and the following use of those proteins for the number of phage displayed antibodies. The CaM-tag permitted the purification of recombinant protein to >90% purity in a single step at yields of >20 mg/l.
Utilizing platelet glycoprotein VI (GP6) as a mannequin, we demonstrated that the recombinant CaM-tagged protein was post-translationally N-glycosylated and had similar ligand specificity to native protein. A novel choice technique, exploiting the CaM tag, was then used to isolate 4 single chain Fv fragments (scFvs) particular for GP6 from a non-immune phage show library. In distinction to different choice strategies, which may end up in antibodies that don’t recognise native protein, the entire scFvs we chosen sure cell floor expressed GP6.
In conclusion, the manufacturing of CaM-tagged proteins in Drosophila Schneider S2 cells and the choice technique reported right here supply benefits over beforehand printed strategies, together with easy tradition circumstances, fast protein purification, particular elution of phage antibodies and preferential number of phage antibodies that recognise native, cell floor expressed protein.
Immunohistochemical examine of Ca2+/calmodulin-dependent protein kinase II within the Drosophila mind utilizing a selected monoclonal antibody.
To investigate the distribution of Drosophila calcium/calmodulin-dependent protein kinase II (dCaMKII) within the grownup mind, we generated monoclonal antibodies in opposition to the bacterially expressed 490-amino acid (a.a.) type of dCaMKII. A type of, named #18 antibody, was used for this examine.
Western blot evaluation of the grownup head extracts confirmed that the antibody particularly detects a number of bands between 55 and 60 kDa comparable to the molecular weights of the splicing isoforms of dCaMKII. Epitope mapping revealed that it was within the area between 199 and 283 a.a. of dCaMKII.
Preferential dCaMKII immunoreactivity within the embryonic nervous system, grownup thoracic ganglion and intestine, and larval neuro-muscular junction (NMJ) was according to earlier observations by in situ hybridization and immunostaining with a polyclonal antibody on the NMJ, indicating that the antibody is relevant to immunohistochemistry.
Though dCaMKII immunoreactive sign was low within the retina, it was discovered at common intervals within the outer margin of the compound eye. These alerts had been most certainly to be interommatidial bristle mechanosensory neurons. dCaMKII immunoreactivity within the mind was noticed in nearly all areas and comparatively increased staining was discovered within the neuropilar area than within the cortex.
Increased dCaMKII immunoreactivity within the mushroom physique (MB) was present in your entire gamma lobe together with the heel, and dorsal suggestions of the alpha and alpha’ lobes, whereas cores of alpha and beta lobes had been stained mild. Discovering plentiful dCaMKII accumulation within the gamma lobe instructed that this lobe would possibly particularly require excessive ranges of dCaMKII expression to perform correctly amongst MB lobes.
Detection of anti-elongation issue 2 kinase (calmodulin-dependent protein kinase III) antibodies in sufferers with systemic lupus erythematosus.
Elongation issue 2 kinase (eEF-2K), also referred to as calmodulin-dependent protein kinase III, is a member of the calmodulin-mediated signaling pathway that hyperlinks activation of cell floor receptors to cell division. The exercise of eEF-2K is elevated in lots of human cancers and could also be a legitimate goal for anti-cancer remedy. It is without doubt one of the unconventional eukaryotic protein kinases with respect to its structural domains compared to different members of the serine/threonine protein kinase superfamily. eEF-2K is extremely conserved in nature.
For instance, the amino acid sequence of human eEF-2K is 90% similar to mouse and rat eEF-2Ks and 40% similar to that of the C. elegans enzyme. Subsequently it has been troublesome to generate high-titer and high-specificity antibodies to the human enzyme by conventional strategies. Sufferers with systemic lupus erythematosus (SLE) produce auto-antibodies to quite a lot of mobile proteins, together with members of the protein translation equipment.
 Calmodulin antibody |
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| 10R-7828 | Fitzgerald | 100 ug | EUR 322 |
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Description: Mouse monoclonal Calmodulin antibody |
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Calmodulin antibody |
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| 10R-C108c | Fitzgerald | 100 ug | EUR 728 |
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Description: Mouse monoclonal Calmodulin antibody |
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Calmodulin antibody |
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| 10R-C109a | Fitzgerald | 200 ul | EUR 544 |
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Description: Mouse monoclonal Calmodulin antibody |
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Calmodulin antibody |
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| 10R-C109b | Fitzgerald | 200 ul | EUR 651 |
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Description: Mouse monoclonal Calmodulin antibody |
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 Calmodulin Antibody |
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| 48759-100ul | SAB | 100ul | EUR 333 |
Calmodulin Antibody |
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| 48759-50ul | SAB | 50ul | EUR 239 |
Calmodulin antibody |
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| 70R-49445 | Fitzgerald | 100 ul | EUR 244 |
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Description: Purified Polyclonal Calmodulin antibody |
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Calmodulin antibody |
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| 70R-33451 | Fitzgerald | 100 ug | EUR 327 |
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Description: Rabbit polyclonal Calmodulin antibody |
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Calmodulin Antibody |
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| AF6353 | Affbiotech | 200ul | EUR 304 |
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Description: Calmodulin Antibody detects endogenous levels of total Calmodulin. |
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Calmodulin Antibody |
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| ABF6353 | Lifescience Market | 100 ug | EUR 438 |
 Calmodulin 2 antibody |
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| 70R-13595 | Fitzgerald | 100 ul | EUR 457 |
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Description: Affinity purified Rabbit polyclonal Calmodulin 2 antibody |
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) Calmodulin antibody (HRP) |
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| 60R-1251 | Fitzgerald | 100 ug | EUR 327 |
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Description: Rabbit polyclonal Calmodulin antibody (HRP) |
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) Calmodulin antibody (FITC) |
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| 60R-1252 | Fitzgerald | 100 ug | EUR 327 |
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Description: Rabbit polyclonal Calmodulin antibody (FITC) |
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) Calmodulin antibody (biotin) |
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| 60R-1253 | Fitzgerald | 100 ug | EUR 327 |
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Description: Rabbit polyclonal Calmodulin antibody (biotin) |
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 Calmodulin Polyclonal Antibody |
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| 42501-100ul | SAB | 100ul | EUR 333 |
Calmodulin Polyclonal Antibody |
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| 40668-100ul | SAB | 100ul | EUR 252 |
Calmodulin Polyclonal Antibody |
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| 40668-50ul | SAB | 50ul | EUR 187 |
 Antibody) Calmodulin (CALM2) Antibody |
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| 20-abx009522 | Abbexa |
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Calmodulin (CALM2) Antibody |
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| abx010483-100ug | Abbexa | 100 ug | EUR 439 |
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Calmodulin (CALM2) Antibody |
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| abx025034-400ul | Abbexa | 400 ul | EUR 523 |
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Calmodulin (CALM2) Antibody |
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| abx025034-80l | Abbexa | 80 µl | EUR 286 |
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Calmodulin (CALM2) Antibody |
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| 20-abx006054 | Abbexa |
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 Antibody) Calmodulin (CAM) Antibody |
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| 20-abx175677 | Abbexa |
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Calmodulin (CAM) Antibody |
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| 20-abx100308 | Abbexa |
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Calmodulin (CALM2) Antibody |
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| 20-abx100309 | Abbexa |
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Calmodulin (CALM2) Antibody |
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| 20-abx100310 | Abbexa |
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) Calmodulin Antibody (Biotin) |
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| 20-abx105021 | Abbexa |
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Calmodulin Antibody (Biotin) |
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| 20-abx105022 | Abbexa |
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Calmodulin Antibody (Biotin) |
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| 20-abx105205 | Abbexa |
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) Calmodulin Antibody (FITC) |
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| 20-abx106435 | Abbexa |
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Calmodulin Antibody (FITC) |
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| 20-abx106622 | Abbexa |
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Therefore, we developed an ELISA assay that would detect anti-eEF2K antibodies from sera of SLE sufferers utilizing purified eEF-2K as an antigen. We screened 117 sera from SLE sufferers. Excessive-titer anti-eEF-2K antibodies had been detected in 72 topics. One of many high-titer sera was used for additional characterization. The auto-antibody acknowledged eEF-2K on immunoblots and immunoprecipitated the kinase with intact enzyme exercise. In conclusion, anti-eEF-2K antibodies are present in sera of SLE sufferers and are helpful instruments to check the function of this extremely conserved enzyme.
