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GAS41 Colorimetric Cell-Based ELISA

SKU:
CBCAB00965
Product Type:
ELISA Kit
ELISA Type:
Cell Based
Research Area:
Developmental Biology
Reactivity:
Human
Mouse
Detection Method:
Colorimetric
€599
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Description

system_update_altDatasheet

GAS41 Colorimetric Cell-Based ELISA

The GAS41 Colorimetric Cell-Based ELISA Kit is an innovative assay designed for the precise measurement of GAS41 levels in cell lysates and tissue samples. This kit offers exceptional sensitivity and accuracy, allowing for reliable and reproducible results in various research applications.GAS41 is a key component of the chromatin remodeling complex and plays a vital role in gene regulation and cell cycle progression. Dysregulation of GAS41 has been implicated in various diseases, including cancer and neurodegenerative disorders, making it an important target for research and therapeutic development.

With its user-friendly protocol and high-performance capabilities, the GAS41 Colorimetric Cell-Based ELISA Kit is an essential tool for researchers seeking to understand the role of GAS41 in health and disease. Order now and take your research to the next level with this advanced assay.

Product Name:GAS41 Colorimetric Cell-Based ELISA
Product Code:CBCAB00965
ELISA Type:Cell-Based
Target:GAS41
Reactivity:Human, Mouse
Dynamic Range:> 5000 Cells
Detection Method:Colorimetric 450 nmStorage/Stability:4°C/6 Months
Format:96-Well Microplate

The GAS41 Colorimetric Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can detect GAS41 protein expression profile in cells. The kit can be used for measuring the relative amounts of GAS41 in cultured cells as well as screening for the effects that various treatments, inhibitors (ie siRNA or chemicals), or activators have on GAS41.

Qualitative determination of GAS41 concentration is achieved by an indirect ELISA format. In essence, GAS41 is captured by GAS41-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: 8089, UniProt ID: O95619, OMIM: 602116, Unigene: Hs.4029
Gene Symbol:YEATS4
Sub Type:None
UniProt Protein Function:YEATS4: Component of the NuA4 histone acetyltransferase (HAT) complex which is involved in transcriptional activation of select genes principally by acetylation of nucleosomal histones H4 and H2A. This modification may both alter nucleosome - DNA interactions and promote interaction of the modified histones with other proteins which positively regulate transcription. This complex may be required for the activation of transcriptional programs associated with oncogene and proto-oncogene mediated growth induction, tumor suppressor mediated growth arrest and replicative senescence, apoptosis, and DNA repair. NuA4 may also play a direct role in DNA repair when recruited to sites of DNA damage.
UniProt Protein Details:

Protein type:Transcription factor

Chromosomal Location of Human Ortholog: 12q13-q15

Cellular Component: nucleoplasm; NuA4 histone acetyltransferase complex; nuclear matrix; cytoplasm; nucleolus

Molecular Function:protein C-terminus binding; protein binding; DNA binding; structural constituent of cytoskeleton; transcription factor activity

Biological Process: mitosis; establishment and/or maintenance of chromatin architecture; transcription, DNA-dependent; regulation of growth; positive regulation of transcription, DNA-dependent

NCBI Summary:The protein encoded by this gene is found in the nucleoli. It has high sequence homology to human MLLT1, and yeast and human MLLT3 proteins. Both MLLT1 and MLLT3 proteins belong to a class of transcription factors, indicating that the encoded protein might also represent a transcription factor. This protein is thought to be required for RNA transcription. This gene has been shown to be amplified in tumors. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jul 2014]
UniProt Code:O95619
NCBI GenInfo Identifier:59799075
NCBI Gene ID:8089
NCBI Accession:O95619.1
UniProt Secondary Accession:O95619,Q9NQD0,
UniProt Related Accession:O95619
Molecular Weight:227
NCBI Full Name:YEATS domain-containing protein 4
NCBI Synonym Full Names:YEATS domain containing 4
NCBI Official Symbol:YEATS4  
NCBI Official Synonym Symbols:YAF9; GAS41; NUBI-1; 4930573H17Rik; B230215M10Rik  
NCBI Protein Information:YEATS domain-containing protein 4; nuBI1; NuMA binding protein 1; nuMA-binding protein 1; glioma-amplified sequence 41; glioma-amplified sequence-41
UniProt Protein Name:YEATS domain-containing protein 4
UniProt Synonym Protein Names:Glioma-amplified sequence 41; Gas41; NuMA-binding protein 1; NuBI-1; NuBI1
Protein Family:YEATS domain-containing protein
UniProt Gene Name:YEATS4  
UniProt Entry Name:YETS4_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-GAS41 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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