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Synapsin1 Colorimetric Cell-Based ELISA Kit

SKU:
CBCAB00366
Product Type:
ELISA Kit
ELISA Type:
Cell Based
Reactivity:
Human
Mouse
Rat
Detection Method:
Colorimetric
€599
Frequently bought together:

Description

system_update_altDatasheetMSDS

Synapsin1 Colorimetric Cell-Based ELISA Kit

The Synapsin1 Colorimetric Cell-Based ELISA Kit from Assay Genie is a powerful tool for the accurate measurement of Synapsin1 levels in cell lysates and tissue homogenates. This kit boasts high sensitivity and specificity, providing reliable and reproducible results for a variety of research applications.Synapsin1 is a key protein involved in synaptic function and neuronal communication, playing a crucial role in conditions such as Alzheimer's disease, Parkinson's disease, and other neurodegenerative disorders.

By measuring Synapsin1 levels, researchers can gain valuable insights into the pathophysiology of these diseases and potential therapeutic targets.Overall, the Synapsin1 Colorimetric Cell-Based ELISA Kit is a valuable asset for researchers studying neurological disorders and looking to unravel the complexities of synaptic transmission. Get accurate and reliable results with this innovative kit from Assay Genie.

Product Name:Synapsin1 Colorimetric Cell-Based ELISA
Product Code:CBCAB00366
ELISA Type:Cell-Based
Target:Synapsin1
Reactivity:Human, Mouse, Rat
Dynamic Range:> 5000 Cells
Detection Method:Colorimetric 450 nmStorage/Stability:4°C/6 Months
Format:96-Well Microplate

The Synapsin1 Colorimetric Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can detect Synapsin1 protein expression profile in cells. The kit can be used for measuring the relative amounts of Synapsin1 in cultured cells as well as screening for the effects that various treatments, inhibitors (ie siRNA or chemicals), or activators have on Synapsin1.

Qualitative determination of Synapsin1 concentration is achieved by an indirect ELISA format. In essence, Synapsin1 is captured by Synapsin1-specific primary antibodies while the HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Through this binding, the HRP enzyme conjugated to the secondary antibody can catalyze a colorimetric reaction upon substrate addition. Due to the qualitative nature of the Cell-Based ELISA, multiple normalization methods are needed:

1. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target absorbance values.
2. Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method may be used to determine cell density. After staining, the results can be analysed by normalizing the absorbance values to cell amounts, by which the plating difference can be adjusted.
Database Information:Gene ID: 6853, UniProt ID: P17600, OMIM: 300491/313440, Unigene: Hs.225936
Gene Symbol:SYN1
Sub Type:None
UniProt Protein Function:SYN1: neuronal phosphoprotein which associates with the cytoplasmic surface of synaptic vesicles and binds to the cytoskeleton. May function in the regulation of neurotransmitter release and of axonogenesis and synaptogenesis. Mutations may be associated with X-linked disorders with primary neuronal degeneration such as Rett syndrome. Two differentially spiced isoforms have been reported.
UniProt Protein Details:

Protein type:Vesicle

Chromosomal Location of Human Ortholog: Xp11.23

Cellular Component: synaptic vesicle

Molecular Function:ATP binding; protein binding; protein kinase binding; transporter activity

Biological Process: regulation of neurotransmitter secretion; synaptic transmission

Disease: Epilepsy, X-linked, With Variable Learning Disabilities And Behavior Disorders

NCBI Summary:This gene is a member of the synapsin gene family. Synapsins encode neuronal phosphoproteins which associate with the cytoplasmic surface of synaptic vesicles. Family members are characterized by common protein domains, and they are implicated in synaptogenesis and the modulation of neurotransmitter release, suggesting a potential role in several neuropsychiatric diseases. This member of the synapsin family plays a role in regulation of axonogenesis and synaptogenesis. The protein encoded serves as a substrate for several different protein kinases and phosphorylation may function in the regulation of this protein in the nerve terminal. Mutations in this gene may be associated with X-linked disorders with primary neuronal degeneration such as Rett syndrome. Alternatively spliced transcript variants encoding different isoforms have been identified. [provided by RefSeq, Jul 2008]
UniProt Code:P17600
NCBI GenInfo Identifier:73920800
NCBI Gene ID:6853
NCBI Accession:P17600.3
UniProt Secondary Accession:P17600,O75825, Q5H9A9, B1AJQ1,
UniProt Related Accession:P17600
Molecular Weight:70,033 Da
NCBI Full Name:Synapsin-1
NCBI Synonym Full Names:synapsin I
NCBI Official Symbol:SYN1  
NCBI Official Synonym Symbols:SYNI; SYN1a; SYN1b  
NCBI Protein Information:synapsin-1
UniProt Protein Name:Synapsin-1
UniProt Synonym Protein Names:Brain protein 4.1; Synapsin I
Protein Family:Synapsin
UniProt Gene Name:SYN1  
UniProt Entry Name:SYN1_HUMAN
ComponentQuantity
96-Well Cell Culture Clear-Bottom Microplate2 plates
10X TBS24 mL
Quenching Buffer24 mL
Blocking Buffer50 mL
15X Wash Buffer50 mL
Primary Antibody Diluent12 mL
100x Anti-Phospho Target Antibody 60 µL
100x Anti-Target Antibody60 µL
Anti-GAPDH Antibody60 µL
HRP-Conjugated Anti-Rabbit IgG Antibody12 mL
HRP-Conjugated Anti-Mouse IgG Antibody12 mL
SDS Solution12 mL
Stop Solution24 mL
Ready-to-Use Substrate12 mL
Crystal Violet Solution12 mL
Adhesive Plate Seals2 seals

The following materials and/or equipment are NOT provided in this kit but are necessary to successfully conduct the experiment:

  • Microplate reader able to measure absorbance at 450 nm and/or 595 nm for Crystal Violet Cell Staining (Optional)
  • Micropipettes with capability of measuring volumes ranging from 1 µL to 1 ml
  • 37% formaldehyde (Sigma Cat# F-8775) or formaldehyde from other sources
  • Squirt bottle, manifold dispenser, multichannel pipette reservoir or automated microplate washer
  • Graph paper or computer software capable of generating or displaying logarithmic functions
  • Absorbent papers or vacuum aspirator
  • Test tubes or microfuge tubes capable of storing ≥1 ml
  • Poly-L-Lysine (Sigma Cat# P4832 for suspension cells)
  • Orbital shaker (optional)
  • Deionized or sterile water

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

StepProcedure
1.Seed 200 µL of 20,000 adherent cells in culture medium in each well of a 96-well plate. The plates included in the kit are sterile and treated for cell culture. For suspension cells and loosely attached cells, coat the plates with 100 µL of 10 µg/ml Poly-L-Lysine (not included) to each well of a 96-well plate for 30 minutes at 37°C prior to adding cells.
2.Incubate the cells for overnight at 37°C, 5% CO2.
3.Treat the cells as desired.
4.Remove the cell culture medium and rinse with 200 µL of 1x TBS, twice.
5.Fix the cells by incubating with 100 µL of Fixing Solution for 20 minutes at room temperature. The 4% formaldehyde is used for adherent cells and 8% formaldehyde is used for suspension cells and loosely attached cells.
6.Remove the Fixing Solution and wash the plate 3 times with 200 µL 1x Wash Buffer for five minutes each time with gentle shaking on the orbital shaker. The plate can be stored at 4°C for a week.
7.Add 100 µL of Quenching Buffer and incubate for 20 minutes at room temperature.
8.Wash the plate 3 times with 1x Wash Buffer for 5 minutes each time.
9.Add 200 µL of Blocking Buffer and incubate for 1 hour at room temperature.
10. Wash 3 times with 200 µL of 1x Wash Buffer for 5 minutes each time.
11.Add 50 µL of 1x primary antibodies (Anti-Synapsin1 Antibody and/or Anti-GAPDH Antibody) to the corresponding wells, cover with Parafilm and incubate for 16 hours (overnight) at 4°C. If the target expression is known to be high, incubate for 2 hours at room temperature.
12.Wash 3 times with 200 µL of 1x Wash Buffer for 5 minutes each time.
13.Add 50 µL of 1x secondary antibodies (HRP-Conjugated AntiRabbit IgG Antibody or HRP-Conjugated Anti-Mouse IgG Antibody) to corresponding wells and incubate for 1.5 hours at room temperature.
14.Wash 3 times with 200 µL of 1x Wash Buffer for 5 minutes each time.
15.Add 50 µL of Ready-to-Use Substrate to each well and incubate for 30 minutes at room temperature in the dark.
16.Add 50 µL of Stop Solution to each well and read OD at 450 nm immediately using the microplate reader.

(Additional Crystal Violet staining may be performed if desired – details of this may be found in the kit technical manual.)

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