A-6442

antibody from Invitrogen Antibodies

Targeting: SYN1 MRX50

Provider product page for A-6442

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [42]
Comments [0]
Validations
Immunocytochemistry [1]
Immunohistochemistry [1]
Other assay [21]

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Product number: A-6442 - Provider product page
Provider: Invitrogen Antibodies
Product name: Synapsin 1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Purifed from natural sources
Description: This antibody has little or no activity against Synapsin II.
Concentration: 0.2 mg/mL

SYN1 protein structure - A-6442 shown in red.

Supportive validation

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Supportive validation

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Supportive validation

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Supportive validation

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Supportive validation

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Supportive validation

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Supportive validation

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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

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Experimental details: NULL

Supportive validation

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Supportive validation

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Experimental details: Figure 1 Marker expression of NTERA2 (NT2) cells and ATRA differentiated NTERA2 neurons (NT2N). NT2 cells (A-C) co-express GFAP and SSEA3 with GFAP staining confined to the nucleus (A). NT2 cells also co-express GFAP and beta-tubulin III (BT3) which are both mainly confined to the nucleus (B). Nestin with GFAP are co-localized in NT2 cells (C). NT2N assembled themselves into cell clusters which co-expressed GFAP and BT3 found within the cytoplasm and neurites (D). Arrow heads point to cells that appear as radial glia cells which only display GFAP staining (D). NT2N also express neural markers TAU and MAP2 with MAP2 confined to dendrites (tapering morphology shown by arrow heads) and TAU not displaying restriction to axons. In merge panel insert shows close-up view of neurite (E). Synapsin 1 staining is seen as punctate along neurite projections co-expressing BT3 (F). Photomicrographs were obtained at 100X (A-C; scale bar 200 um) and 200X (D-F; scale bar 100 um).

Supportive validation

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Experimental details: Figure 5 Dendritic and synaptic proteins in the aged human hippocampus: Differential association with CAm. Immunoperoxidase-hematoxylin staining of brain sections obtained from an 85 year-old ND individual. A) Representative images of the CA2 subfield and fornix (A') labeled with the dendritic cytoskeletal marker anti-MAP2. Note the strong immunoreactivity in pyramidal cell bodies and dendrites, as well as CAm (A') . B- C) Immunoperoxidase staining using anti-Synapsin-1 (B) and anti-Synaptophysin antibodies (C) depicting the CA2 pyramidal cell layer (B, C) , the dentate gyrus molecular layer (B', C') and the fornix (B"", C"") . While a characteristic synaptic immunoreactivity of both markers was evident in the hippocampus proper and dentate gyrus, no accumulation of these synaptic proteins was seen in CAm. Scale bars: A- C = 30 mum, A'- C' = 30 mum.

Supportive validation

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Experimental details: Figure 4 The synaptic potential of 21- and 28-day-old hiPSC- and hESC-derived neurons. (a-b) In the denervated cochlear explant controls at 1 DIV, some accumulation of residual NFM within hair cell somata and a few synaptic puncta were observed. (c-d) After 10 DIV, there appeared to be fewer hair cells with NFM accumulation and synapsin 1 was undetectable. (e) Light microscope image of denervated cochlear explant cocultured with stem cell-derived neurosphere. (f and g) The hiPSC and hESC neurospheres cocultured at 21 DIV extended their neural processes towards hair cells in the denervated explants. Punctate-like synapsin 1 expression was observed along the neural processes of both the hiPSC- and hESC-derived neurites making contact with hair cells. (h and i) The hiPSC and hESC neurospheres cocultured at 28 DIV projected fewer neural processes towards hair cells in the denervated explants. Synapsin 1 was rarely observed along the neural processes of both the hiPSC- and hESC-derived neurites making contact with hair cells. The merged 1 images represent higher magnification images of the boxed inserts and depict the punctate-like synaptic contacts made between the stem cell-derived neural processes and hair cells. Scale bar = 20 mu m (applicable for all images). (j) The 21-day-old hiPSC- and hESC-derived neurons made contact with a significantly higher number of hair cells, compared to the 28-day-old neurons (H9: p < 0.05). Furthermore, the 21-day-old hESC neurons made contact w

Supportive validation

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Experimental details: Figure 3 Depletion of TDP-43 from Microglia Enhances Amyloid Clearance but Exacerbates Synaptic Loss in a Mouse Model of AD (A) Multiplexed electrocheminoluminescent assay measurements of Abeta40 and sAPPbeta levels in the SDS-soluble fraction of cortex homogenates from 7-month-old APP arc mice lacking TDP-43 in microglia. Mean +- SEM, n = 4 mice per genotype, ** p < 0.01, using two-way ANOVA, followed by Sidak's post hoc test. (B) Representative max-projections of confocal stacks from cortex of WT;APP arc or cKO;APP arc mice stained with Thioflavin S. (C and D) Quantification of ThioS plaque density (C) and area covered by plaques from the cortex of 7-month-old APP arc , WT (n = 36) and cKO (n = 36) with acquisitions from 4 animals per genotype (D). Mean +- SEM, ** p < 0.01, using two-tailed unpaired t test. (E-I) Representative blots for synaptic markers, from the cortex of 7-month-old APP arc mice, WT or KO for microglial TDP-43 (E). Quantification of western blots for PSD95 (F), MAP2 (G), synapsin (H), and synaptophysin (I) normalized for GAPDH reference gene. Mean +- SEM, n = 4-5 mice per genotype, * p < 0.05, ** p < 0.01, using two-tailed unpaired t test. (J and K), Representative 3D reconstruction from confocal acquisitions of vGlut1 immunoreactivity in the cortex of WT and cKO mice (J) and relative quantification (K) (WT n = 25, cKO n = 17, acquisitions from 4 mice per genotype; **** p < 0.0001, two-tailed t test).

Supportive validation

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Experimental details: Comparative immunoreactivity of a selected panel of antibodies in perfusion-fixed and immersion-fixed tissue. Immersion fixation was carried out immediately following perfusion with ACSF (see ). Images are from immunoperoxidase staining and photographed in bright-field (A-F; G', H', I', J') or dark-field (G, H, I, J) transmission microscopy. Each staining pattern is illustrated at two different magnifications to highlight the regional and cellular distribution of the markers investigated. The regions selected include the dorsal hippocampus (A-F; I-J) and the occipital cortex (G-H). (A, B) CD 68 is a microglial cell marker, expressed at low levels in resting microglia; as seen in A' and B', both fixation methods are compatible with the staining of fine ramified processes of resting microglia. (C, D) GFAP is a marker of astrocytes, strongly expressed in the hippocampal formation, depicted in tissue from old mice (16 and 19 months) to illustrate that immersion-fixation is also suitable for tissue from aged mice. (E, F) S ynapsin 1 is a presynaptic protein, strongly expressed in mossy fibers innervating CA 3 pyramidal cells; the two fixation protocols provide similar results, with a clear distinction of these terminal in the stratum lucidum (sl) and a weaker staining in the stratum pyramidale (sp) and radiatum (sr). (G, H) Distribution of serotonergic (5- HT ) axons in the neocortex, depicted in both bright- and dark-field to illustrate that a similar signal-to-noise ratio and mo

Supportive validation

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Experimental details: Fig. 2 The silencing efficiency of UBE3C in BrCa cells. a , b The silencing efficiency of UBE3C in BrCa cells was confirmed by qPCR and western blotting analysis. SiRNA #2 and siRNA #1 showed highest silencing efficiency for UBE3C in MCF-7 and MDA-MB-453 cells, respectively. ***P < 0.001. c , d The silencing efficiency of UBE3C in BrCa cells and cytomorphology was determined by microscopy and immunofluorescence analysis. Green fluorescence indicated that the cells have been successfully transfected. ***P < 0.001. Bar = 200 mum

Supportive validation

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Experimental details: Fig. 4 The efficiency of UBE3C overexpression in BrCa cells. a , b The efficiency of UBE3C overexpression in BrCa cells was confirmed by qPCR and western blotting analysis. UBE3C-overexpressing vector showed satisfactory transfection efficiency. c , d The efficiency of UBE3C overexpression in BrCa cells and cytomorphology was determined by microscopy and immunofluorescence analysis. Green fluorescence indicated that the cells have been successfully transfected. ***P < 0.001. Bar = 200 mum

Supportive validation

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Experimental details: Fig. 6 UBE3C promotes the nuclear accumulation of beta-catenin. a , b Immunofluorescence staining shows that silencing or overexpressing UBE3C in BrCa cells inhibited or increased the nuclear accumulation of beta-catenin, respectively. Blue: DAPI, green: beta-catenin, red: UBE3C. Bar = 20 mum. c , d The expression levels of total and nuclear beta-catenin in UBE3C-silenced and/or UBE3C-overexpressing BrCa cells was evaluated by western blotting. UBE3C-silenced BrCa cells showed low nuclear beta-catenin level, but UBE3C-overexpressing BrCa cells were opposite. **P < 0.01, ***P < 0.001

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7. Intraneuronal effect of rHsCTSD on SNCA and synaptical vesicles in primary neurons and in DA-iPSn. (A) Representative immunostaining of neuronal processes of primary neurons derived from WT, ctsd KO and ctsd KO +rHsCTSD animals. SNCA shown by antibody anti-syn-1 in red and SYN-1 (synapsin I) in green. Scale bar: 5 um. (B) Quantification of SYN-1-positive vesicle cluster number per um and (C) of the average SYN-1-positive vesicle size per punctum (um) in primary neurons from WT, ctsd KO and ctsd KO +rHsCTSD (analysis of n = 11-13 individual neurons per group). (D) Representative immunostaining of SNCA (LB509; red) and SYN-1 (green) in neuronal processes of DA-iPSn of isogenic control (A53T corr) and mutant line (A53T) with and without rHsCTSD treatment. Scale bar: 5 um. (E) Quantification of SYN-1-positive vesicle cluster numbers per um and (F) analyses of average SYN-1-positive vesicle size per punctum (um) in neuronal processes of DA-iPSn A53T corr. +PBS, A53T +PBS and A53T +rHsCTSD (n = 13-16 individual neurons per group). (G and H) Representative structured illumination microscopy (SIM) images providing detailed localization of SNCA respective to SYN-1 signal in (G) primary neurons derived from KO +PBS and KO +rHsCTSD animals and (H) DA-iPSn from A53T mutant +PBS and +rHsCTSD. Neuronal processes were immunostained with SNCA (Antibody: syn-1 for primary neurons and LB509 for DA-iPSn; magenta) and SYN-1 (green). Scale bar: overview 2 um and inset 1 um. Images highl