MA5-16402

antibody from Invitrogen Antibodies

Targeting: SYP MRX96

Provider product page for MA5-16402

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [41]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [1]
Immunohistochemistry [1]
Other assay [5]

Submit

Product number: MA5-16402 - Provider product page
Provider: Invitrogen Antibodies
Product name: Synaptophysin Monoclonal Antibody (SP11)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 30 min at room temperature. A suggested positive control is pancreas or neuroendocrine tumor.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Antibody clone number: SP11
Vial size: 500 µL
Concentration: Conc. Not Determined
Storage: -20°C or -80°C if preferred

SYP protein structure - MA5-16402 shown in red.

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure EV3 Synaptophysin and PSD -95 remain unaltered in neurons containing GA aggregates A Primary cortical neurons were co-transfected with mCherry or mCherry-GA n and GCaMP6f. After 48 h, mCherry-positive cells were determined. Green fluorescence intensity was then recorded from identified neurons. Basal fluorescence was monitored prior to induced depolarization via perfusion with ACSF containing 50 mM KCl. Graphical representation is the quantification of peak change in fluorescence (DeltaF) following ACSF perfusion normalized to basal fluorescence (F), DeltaF/F. A significant increase was observed in mCherry-GA n containing cells (GA 25 , GA 50 , GA 200 ) (* P < 0.05); however, a length-dependent association with Ca 2+ influx levels was not detected. Data presented as mean +- SEM. One-way ANOVA, post hoc Dunnett''s multiple comparison test. A total of 8-25 cells per condition were pooled from n = 5 independent biological replicates. B, C eGFP-GA n dipeptides were expressed in cortical or motor neurons for 48 h and then immunostained for neurofilament and either synaptophysin or PSD-95. Z-stack confocal images were captured at 60x magnification; puncta were quantified by ImageJ through manual counting. (B) Quantification of synaptophysin (left) or PSD-95 (right) puncta along neurites in eGFP-GA n expressing cortical neurons compared with eGFP-only expressing cells revealed no significant differences. (C) Quantification of synaptophysin (left) or PSD-95 (right) puncta alon

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 SV 2 is reduced in i3 neurons from a C9orf72- ALS patient A, B i3 cortical neurons from a C9orf72-ALS patient and the paired isogenic control were differentiated for 21 days then were immunostained for neurofilament (red), and SV2 (green) (A) or synaptophysin (green) (B). Shown are representative z-stack confocal images of neurites. C, D Quantification of SV2 (C) or synaptophysin (D) colocalized with neurofilament normalized to neurofilament area. This analysis revealed a significant reduction in SV2 levels in the C9orf72 line compared with the isogenic control, while synaptophysin remained unaltered. Data presented as mean +- SEM. Unpaired t -test, * P < 0.05. 15 non-overlapping regions from three differentiated wells for each genotype were evaluated. E mRNA was Trizol extracted from three independent wells of C9-ALS and isogenic i3 cortical neurons. RNA was converted to cDNA using the Superscript First-strand kit, and qPCR was performed using PowerUp SYBR Green. Measurements were normalized to the housekeeping gene GAPDH and then to isogenic levels. Analysis was performed using the DeltaDeltaCT method. 2 DeltaDeltaCT +- SE is presented. Unpaired t -test, ** P < 0.01. F Whole-cell lysates were generated from three independent wells of C9-ALS and isogenic i3 cortical neurons differentiated for 21 days. Lysates were immunoblotted for SV2 and normalized to total protein loading. Quantification of band intensities revealed a significant decrease in SV2 protein levels in

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 9 SV 2 is specifically reduced in spinal cord lysates of symptomatic transgenic GA 149- CFP mice Spinal cord lysates were generated from fresh-frozen tissue of 20-month-old mice either expressing GA 149 -CFP under the Thy1 promoter (Tgx GA 149 ) or wild-type controls. A-C Lysates were immunoblotted for SV2 (A), synaptophysin (B), and PSD-95 (C). Total protein levels were determined by imaging the Mini-PROTEAN TGX stain-free gel prior to transfer. Quantification of band intensities normalized to total protein loading revealed a significant reduction in SV2 protein levels in the GA-expressing cohort compared with wild-type animals, while synaptophysin and PSD-95 levels remained unaltered. Data presented as mean +- SEM. Unpaired t -test, * P < 0.05. A total of four animals per group were examined. Exact P- value can be found in Appendix Table S1 . Source data are available online for this figure.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 10 SV 2 is specifically reduced at neuromuscular junctions of symptomatic transgenic GA 149 - CFP mice Muscle sections (30 mum) were cut from snap-frozen tissue of 20-month-old GA 149 mice (Tgx GA 149 ) or wild-type controls. Immunohistochemical staining was performed to assess synaptic protein levels in proximity to NMJs. Sections were immunostained for SV2 or synaptophysin and counterstained with neurofilament. A Left Panels: Representative images show SV2 levels (green) in proximity to alpha-bungarotoxin-positive NMJs (red) in wild-type animals (top) versus GA 149 animals (bottom). Right panels: Representative images show SV2 levels (green) overlapping with neurofilament (blue) at synaptic terminals approaching muscle in wild-type animals (top) versus GA 149 animals (bottom). B, C Quantification of SV2 (B) or synaptophysin (C) colocalized with neurofilament and normalized to neurofilament area. This analysis revealed a significant reduction in SV2 levels in GA 149 animals compared with wild types, while synaptophysin remained unaltered. Data presented as mean +- SEM. Unpaired t -test, * P < 0.05. A total of 10 non-overlapping regions of muscle from each of five animals per group were evaluated. Exact P- values can be found in Appendix Table S1 .