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CSE1L Colorimetric Cell-Based ELISA Kit (CBCAB00602)

SKU:
CBCAB00602
Product Type:
ELISA Kit
ELISA Type:
Cell Based
Research Area:
Cell Biology
Reactivity:
Human
Detection Method:
Colorimetric
€769

Description

system_update_altDatasheetMSDS

CSE1L Colorimetric Cell-Based ELISA Kit

The CSE1L Colorimetric Cell-Based ELISA Kit is specifically designed for the easy and accurate detection of Cell Division Cycle Protein 1 homolog (CSE1L) levels in cell lysates. This kit offers high sensitivity and specificity, allowing for reliable and reproducible results in a variety of research applications.CSE1L is a key protein involved in regulating cell division and proliferation, making it an important target for studying cellular processes and diseases such as cancer. By accurately measuring CSE1L levels in cell samples, researchers can gain valuable insights into the mechanisms of cell growth and potential therapeutic interventions.

The CSE1L Colorimetric Cell-Based ELISA Kit is user-friendly and efficient, providing researchers with a powerful tool for advancing their research in cell biology and disease pathology. Whether investigating basic cellular functions or searching for novel therapeutic targets, this kit offers the reliability and precision needed for successful experimentation.

Product Name:CSE1L Colorimetric Cell-Based ELISA
Product Code:CBCAB00602
ELISA Type:Cell-Based
Target:CSE1L
Reactivity:Human
Dynamic Range:> 5000 Cells
Detection Method:Colorimetric 450 nmStorage/Stability:4°C/6 Months
Format:96-Well Microplate

The CSE1L Colorimetric Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can detect CSE1L protein expression profile in cells. The kit can be used for measuring the relative amounts of CSE1L in cultured cells as well as screening for the effects that various treatments, inhibitors (ie siRNA or chemicals), or activators have on CSE1L.

Qualitative determination of CSE1L concentration is achieved by an indirect ELISA format. In essence, CSE1L is captured by CSE1L-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: 1434, UniProt ID: P55060, OMIM: 601342, Unigene: Hs.90073
Gene Symbol:CSE1L
Sub Type:None
UniProt Protein Function:Exportin-2: Export receptor for importin-alpha. Mediates importin- alpha re-export from the nucleus to the cytoplasm after import substrates (cargos) have been released into the nucleoplasm. In the nucleus binds cooperatively to importin-alpha and to the GTPase Ran in its active GTP-bound form. Docking of this trimeric complex to the nuclear pore complex (NPC) is mediated through binding to nucleoporins. Upon transit of a nuclear export complex into the cytoplasm, disassembling of the complex and hydrolysis of Ran-GTP to Ran-GDP (induced by RANBP1 and RANGAP1, respectively) cause release of the importin-alpha from the export receptor. CSE1L/XPO2 then return to the nuclear compartment and mediate another round of transport. The directionality of nuclear export is thought to be conferred by an asymmetric distribution of the GTP- and GDP-bound forms of Ran between the cytoplasm and nucleus. Belongs to the XPO2/CSE1 family. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Nuclear export; Karyopherin; Nuclear import

Chromosomal Location of Human Ortholog: 20q13

Cellular Component: cytoplasm; cytosol; membrane; nuclear envelope; nucleoplasm; nucleus

Molecular Function:nuclear export signal receptor activity; protein binding

Biological Process: apoptosis; cell proliferation; protein import into nucleus

NCBI Summary:Proteins that carry a nuclear localization signal (NLS) are transported into the nucleus by the importin-alpha/beta heterodimer. Importin-alpha binds the NLS, while importin-beta mediates translocation through the nuclear pore complex. After translocation, RanGTP binds importin-beta and displaces importin-alpha. Importin-alpha must then be returned to the cytoplasm, leaving the NLS protein behind. The protein encoded by this gene binds strongly to NLS-free importin-alpha, and this binding is released in the cytoplasm by the combined action of RANBP1 and RANGAP1. In addition, the encoded protein may play a role both in apoptosis and in cell proliferation. Alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Jan 2012]
UniProt Code:P55060
NCBI GenInfo Identifier:62297557
NCBI Gene ID:1434
NCBI Accession:P55060.3
UniProt Secondary Accession:P55060,O75432, Q32M40, Q9H5B7, Q9NTS0, Q9UP98, Q9UP99 Q9UPA0, A3RLL6, B2R5T4, E1P5Y0, F8W904,
UniProt Related Accession:P55060
Molecular Weight:103,879 Da
NCBI Full Name:Exportin-2
NCBI Synonym Full Names:chromosome segregation 1 like
NCBI Official Symbol:CSE1L  
NCBI Official Synonym Symbols:CAS; CSE1; XPO2  
NCBI Protein Information:exportin-2
UniProt Protein Name:Exportin-2
UniProt Synonym Protein Names:Cellular apoptosis susceptibility protein; Chromosome segregation 1-like protein; Importin-alpha re-exporter
UniProt Gene Name:CSE1L  
UniProt Entry Name:XPO2_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-CSE1L 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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