Immunocytochemistry visualizes the precise spatial location of goal molecules. The most typical technique for ultrastructural immunocytochemistry is the conjugation of nanogold particles to antibodies as probes. Nonetheless, typical nanogold labelling requires time-consuming nanogold probe preparation and ultrathin sectioning of cell/tissue samples.
Right here, we introduce an in situ technique involving nanogold nucleation in immunoenzymatic merchandise on common paraffin/cryostat sections and supply distinctive perception into nanogold improvement underneath hot-humid air circumstances. Nanogold particles have been particularly localized on kidney podocytes to focus on synaptopodin.
Transmission electron microscopy revealed secondary development and self-assembly that may very well be experimentally managed by bovine serum albumin stabilization and phosphate-buffered saline acceleration. Useful retrospective nanogold labelling for gastric H+/Ok+-ATPase was achieved on classic immunoenzymatic deposits after a protracted lapse of 15 years (i.e., 15-year-old deposits).
The current in situ nanogold labelling is anticipated to fill the hole between mild and electron microscopy to correlate cell/tissue construction and performance.
Maternal hypoxia developmentally applications low podocyte endowment in male, however not feminine offspring.
Fetal hypoxia is a typical complication of being pregnant. We have now beforehand reported that maternal hypoxia in late gestation in mice provides rise to male offspring with diminished nephron quantity, whereas females have regular nephron quantity.
Male offspring later develop proteinuria and renal pathology, together with glomerular pathology, whereas feminine offspring are unaffected. Given the central function of podocyte depletion in glomerular and renal pathology, we examined whether or not maternal hypoxia resulted in low podocyte endowment in offspring. Pregnant CD1 mice have been allotted at embryonic day 14.5 to normoxic (21% oxygen) or hypoxic (12% oxygen) circumstances.
At postnatal day 21, kidneys from mice have been immersion fastened, and one mid-hilar slice per kidney was immunostained with antibodies directed in opposition to p57 and synaptopodin for podocyte identification. Slices have been cleared and imaged with a multiphoton microscope for podometric evaluation. Male hypoxic offspring had considerably decrease beginning weight, nephron quantity, and podocyte endowment than normoxic male offspring (podocyte quantity; normoxic 62.86 ± 2.26 podocytes per glomerulus, hypoxic 53.38 ± 2.25; p < .01, imply ± SEM).
In distinction, hypoxic feminine offspring had low beginning weight however their nephron and podocyte endowment was the identical as normoxic feminine offspring (podocyte quantity; normoxic 62.38 ± 1.86 podocytes per glomerulus, hypoxic 61.81 ± 1.80; p = .88).
To the perfect of our data, that is the primary report of developmentally programmed low podocyte endowment. Given the well-known affiliation between podocyte depletion in maturity and glomerular pathology, we postulate that podocyte endowment might place offspring vulnerable to renal illness in maturity, and clarify the better vulnerability of male offspring.
Dynamic Regulation of Synaptopodin and the Axon Preliminary Section in Retinal Ganglion Cells Throughout Postnatal Growth.
A key part permitting a neuron to perform correctly inside its dynamic surroundings is the axon preliminary phase (AIS), the positioning of motion potential era. In visible cortex, AIS of pyramidal neurons bear intervals of activity-dependent structural plasticity throughout improvement.
Nonetheless, it stays unknown how AIS morphology is organized throughout improvement for downstream cells within the visible pathway (retinal ganglion cells; RGCs) and whether or not AIS retain the flexibility to dynamically alter to adjustments in community state. Right here, we investigated the maturation of AIS in RGCs throughout mouse retinal improvement, and examined putative activity-dependent mechanisms by making use of visible deprivation with a give attention to the AIS-specific cisternal organelle (CO), a presumed Ca2+-store.
Complete-mount retinae from wildtype and Thy1-GFP transgenic mice have been processed for multi-channel immunofluorescence utilizing antibodies in opposition to AIS scaffolding proteins ankyrin-G, βIV-spectrin and the CO marker synaptopodin (synpo).
Confocal microscopy together with morphometrical evaluation of AIS size and place in addition to synpo cluster dimension was carried out. Information indicated {that a} subset of RGC AIS comprises synpo clusters and that these present important dynamic regulation in dimension throughout improvement in addition to after visible deprivation.
Utilizing tremendous decision microscopy, we addressed the subcellular localization of synpo in RGC axons. Just like cortical neurons, RGCs present a periodic distribution of AIS scaffolding proteins. A beforehand reported scaffold-deficient nanodomain correlating with synpo localization shouldn’t be evident in all RGC AIS.
In abstract, our work demonstrates a dynamic regulation of each the AIS and synpo in RGCs throughout retinal improvement and after visible deprivation, offering first proof that the AIS and CO in RGCs can bear structural plasticity in response to adjustments in community exercise.
Partial podocyte replenishment in experimental FSGS derives from nonpodocyte sources.
The present research used genetic destiny mapping to show that grownup podocytes might be partially replenished following depletion. Inducible NPHS2-rtTA/tetO-Cre/RS-ZsGreen-R reporter mice have been generated to completely label podocytes with the ZsGreen reporter. Experimental focal segmental glomerulosclerosis (FSGS) was induced with a cytotoxic podocyte antibody.
On FSGS day 7, immunostaining for the podocyte markers p57, synaptopodin, and podocin have been markedly decreased by 44%, and this was accompanied by a lower in ZsGreen fluorescence. The nuclear stain DAPI was absent in segments of diminished ZsGreen and podocyte marker staining, which is per podocyte depletion.
Staining for p57, synaptopodin, podocin, and DAPI elevated at FSGS day 28 and was augmented by the ACE inhibitor enalapril, which is per a partial replenishment of podocytes. In distinction, ZsGreen fluorescence didn’t return and remained considerably low at day 28, indicating replenishment was from a nonpodocyte origin.
Regardless of administration of bromodeoxyuridine (BrdU) thrice weekly all through the course of illness, BrdU staining was not detected in podocytes, which is per an absence of proliferation. Though ZsGreen reporting was diminished within the tuft at FSGS day 28, labeled podocytes have been detected alongside the Bowman’s capsule in a subset of glomeruli, which is per migration from the tuft.
Furthermore, greater than half of the migrated podocytes coexpressed the parietal epithelial cell (PEC) proteins claudin-1, SSeCKS, and PAX8. These outcomes present that though podocytes might be partially replenished following abrupt depletion, a course of augmented by ACE inhibition, the supply or sources are nonpodocyte in origin and are unbiased of proliferation. Moreover, a subset of podocytes migrate to the Bowman’s capsule and start to coexpress PEC markers.
Renin-Angiotensin-Aldosterone System Inhibition Will increase Podocyte Derivation from Cells of Renin Lineage.
As a result of grownup podocytes can’t proliferate and are subsequently unable to self-renew, substitute of those cells is dependent upon stem/progenitor cells. Though podocyte quantity is greater after renin-angiotensin-aldosterone system (RAAS) inhibition in glomerular illnesses, the occasions explaining this enhance are unclear.
Cells of renin lineage (CoRL) have marked plasticity, together with the flexibility to amass a podocyte phenotype. To check the speculation that RAAS inhibition partially replenishes grownup podocytes by growing CoRL quantity, migration, and/or transdifferentiation, we administered tamoxifen to Ren1cCreERxRs-tdTomato-R CoRL reporter mice to induce everlasting labeling of CoRL with purple fluorescent protein variant tdTomato.
We then induced experimental FSGS, typified by abrupt podocyte depletion, with a cytopathic antipodocyte antibody. RAAS inhibition by enalapril (angiotensin-converting enzyme inhibitor) or losartan (angiotensin-receptor blocker) in FSGS mice stimulated the proliferation of CoRL, growing the reservoir of those cells within the juxtaglomerular compartment (JGC).
In contrast with water or hydralazine, RAAS inhibition considerably elevated the migration of CoRL from the JGC to the intraglomerular compartment (IGC), with extra glomeruli containing RFP+CoRL and, inside these glomeruli, extra RFP+CoRL.
Synaptopodin (SYNPO) Antibody |
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| abx020655-005mg | Abbexa | 0.05 mg | EUR 1279.2 |
Synaptopodin (SYNPO) Antibody |
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| abx020657-5ml | Abbexa | 5 ml | EUR 1195.2 |
Synaptopodin (SYNPO) Antibody |
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| abx027982-400ul | Abbexa | 400 ul | EUR 627.6 |
Synaptopodin (SYNPO) Antibody |
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| abx027982-80l | Abbexa | 80 µl | EUR 343.2 |
Synaptopodin (SYNPO) Antibody |
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| 20-abx339139 | Abbexa |
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Synaptopodin (SYNPO) Antibody |
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| 20-abx174689 | Abbexa |
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Synaptopodin (SYNPO) Antibody |
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| abx238423-100ug | Abbexa | 100 ug | EUR 610.8 |
Anti-Synaptopodin antibody |
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| PAab08423 | Lifescience Market | 100 ug | EUR 463.2 |
anti- Synaptopodin antibody |
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| FNab08423 | FN Test | 100µg | EUR 658.5 |
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Description: Antibody raised against Synaptopodin |
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Anti-Synaptopodin antibody |
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| STJ13100352 | St John's Laboratory | 100 µl | EUR 512.4 |
Anti-Synaptopodin antibody |
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| STJ13100378 | St John's Laboratory | 100 µl | EUR 512.4 |
Synaptopodin Antibody / SYNPO |
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| R32281 | NSJ Bioreagents | 100 ug | EUR 419 |
Synaptopodin Antibody / SYNPO |
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| RQ5766 | NSJ Bioreagents | 100 ug | EUR 419 |
Synaptopodin 2 (SYNPO2) Antibody |
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| 20-abx178518 | Abbexa |
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Synaptopodin 2 (SYNPO2) Antibody |
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| 20-abx178519 | Abbexa |
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Synaptopodin 2 (SYNPO2) Antibody |
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| 20-abx178520 | Abbexa |
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Synaptopodin 2 (SYNPO2) Antibody |
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| 20-abx210716 | Abbexa |
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Synaptopodin 2 (SYNPO2) Antibody |
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| 20-abx210717 | Abbexa |
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Synaptopodin 2 (SYNPO2) Antibody |
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| 20-abx174690 | Abbexa |
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Anti-Synaptopodin/SYNPO Antibody |
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| PA1384 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-Synaptopodin/SYNPO Antibody |
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| RP1069 | BosterBio | 100ug/vial | EUR 352.8 |
Synaptopodin Blocking Peptide |
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| DF12173-BP | Affbiotech | 1mg | EUR 234 |
Recombinant Synaptopodin (SYNPO) |
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| 4-RPC885Hu01 | Cloud-Clone |
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Description: Recombinant Human Synaptopodin expressed in: E.coli |
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Recombinant Synaptopodin (SYNPO) |
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| 4-RPC885Hu02 | Cloud-Clone |
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Description: Recombinant Human Synaptopodin expressed in: E.coli |
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Recombinant Synaptopodin (SYNPO) |
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| 4-RPC885Hu03 | Cloud-Clone |
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Description: Recombinant Human Synaptopodin expressed in: E.coli |
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Synaptopodin 2-Like (SYNPO2L) Antibody |
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| 20-abx115944 | Abbexa |
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Synaptopodin 2-Like Protein (SYNPO2L) Antibody |
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| 20-abx002526 | Abbexa |
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Synaptopodin 2-Like Protein (SYNPO2L) Antibody |
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| abx238441-100ug | Abbexa | 100 ug | EUR 610.8 |
Polyclonal SYNPO / Synaptopodin Antibody (C-Terminus) |
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| APR13636G | Leading Biology | 0.05mg | EUR 580.8 |
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Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human SYNPO / Synaptopodin (C-Terminus). This antibody is tested and proven to work in the following applications: |
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Synaptopodin (SYNPO) Polyclonal Antibody (Human, Mouse) |
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| 4-PAC885Hu01 | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Mouse Synaptopodin (SYNPO) |
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Human Synaptopodin (SYNPO) Protein |
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| 20-abx069219 | Abbexa |
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Goat Synaptopodin ELISA kit |
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| E06S0219-192T | BlueGene | 192 tests | EUR 1524 |
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Description: A sandwich ELISA for quantitative measurement of Goat Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Goat Synaptopodin ELISA kit |
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| E06S0219-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
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Description: A sandwich ELISA for quantitative measurement of Goat Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Goat Synaptopodin ELISA kit |
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| E06S0219-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
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Description: A sandwich ELISA for quantitative measurement of Goat Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Rabbit Synaptopodin ELISA kit |
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| E04S0219-192T | BlueGene | 192 tests | EUR 1524 |
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Description: A sandwich ELISA for quantitative measurement of Rabbit Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Rabbit Synaptopodin ELISA kit |
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| E04S0219-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
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Description: A sandwich ELISA for quantitative measurement of Rabbit Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Rabbit Synaptopodin ELISA kit |
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| E04S0219-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
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Description: A sandwich ELISA for quantitative measurement of Rabbit Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Mouse Synaptopodin ELISA kit |
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| E03S0219-192T | BlueGene | 192 tests | EUR 1524 |
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Description: A sandwich ELISA for quantitative measurement of Mouse Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Mouse Synaptopodin ELISA kit |
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| E03S0219-48 | BlueGene | 1 plate of 48 wells | EUR 624 |
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Description: A sandwich ELISA for quantitative measurement of Mouse Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Mouse Synaptopodin ELISA kit |
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| E03S0219-96 | BlueGene | 1 plate of 96 wells | EUR 822 |
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Description: A sandwich ELISA for quantitative measurement of Mouse Synaptopodin in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species. |
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Recombinant human Synaptopodin-2 |
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| P1255 | FN Test | 100ug | Ask for price |
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Description: Recombinant protein for human Synaptopodin-2 |
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Furthermore, RAAS inhibition in FSGS mice elevated RFP+CoRL transdifferentiation within the IGC to phenotypes, per these of podocytes (coexpression of synaptopodin and Wilms tumor protein), parietal epithelial cells (PAX 8), and mesangial cells (α8 integrin). These outcomes present that within the context of podocyte depletion in FSGS, RAAS inhibition augments CoRL proliferation and plasticity towards three completely different glomerular cell lineages.
