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IL-3Rbeta Colorimetric Cell-Based ELISA Kit

SKU:
CBCAB00314
Product Type:
ELISA Kit
ELISA Type:
Cell Based
Reactivity:
Human
Mouse
Rat
Detection Method:
Colorimetric
€599
Frequently bought together:

Description

system_update_altDatasheetMSDS

IL-3Rbeta Colorimetric Cell-Based ELISA Kit

The IL-3Rbeta Colorimetric Cell-Based ELISA Kit is specifically designed for the precise measurement of IL-3 receptor beta levels in cell samples. This innovative kit offers exceptional sensitivity and accuracy, providing researchers with reliable and consistent results for a variety of applications.IL-3 receptor beta is a key protein involved in the regulation of immune responses and hematopoiesis. Its dysregulation has been implicated in various disease processes, including leukemia and autoimmune disorders.

By detecting IL-3 receptor beta levels, researchers can gain valuable insights into the underlying mechanisms of these diseases and potentially identify novel therapeutic targets.With its user-friendly protocol and high-performance capabilities, the IL-3Rbeta Colorimetric Cell-Based ELISA Kit is an essential tool for investigating the role of IL-3 receptor beta in cellular processes and disease pathology. Order now to elevate your research to the next level.

Product Name:IL-3Rbeta Colorimetric Cell-Based ELISA
Product Code:CBCAB00314
ELISA Type:Cell-Based
Target:IL-3Rbeta
Reactivity:Human, Mouse, Rat
Dynamic Range:> 5000 Cells
Detection Method:Colorimetric 450 nmStorage/Stability:4°C/6 Months
Format:96-Well Microplate

The IL-3Rbeta Colorimetric Cell-Based ELISA Kit is a convenient, lysate-free, high throughput and sensitive assay kit that can detect IL-3Rbeta protein expression profile in cells. The kit can be used for measuring the relative amounts of IL-3Rbeta in cultured cells as well as screening for the effects that various treatments, inhibitors (ie siRNA or chemicals), or activators have on IL-3Rbeta.

Qualitative determination of IL-3Rbeta concentration is achieved by an indirect ELISA format. In essence, IL-3Rbeta is captured by IL-3Rbeta-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: 1439, UniProt ID: P32927, OMIM: 138981, Unigene: Hs.592192
Gene Symbol:CSF2RB
Sub Type:None
UniProt Protein Function:CSF2RB: a common beta chain of the high affinity receptor for IL-3, IL-5 and CSF. A type I cytokine family of receptor. Defects are a cause of congenital pulmonary alveolar proteinosis (PAP) [MIM:265120]. PAP is an autosomal recessive fatal respiratory disease.
UniProt Protein Details:

Protein type:Membrane protein, integral; Receptor, cytokine

Chromosomal Location of Human Ortholog: 22q13.1

Cellular Component: granulocyte macrophage colony-stimulating factor receptor complex; plasma membrane

Molecular Function:interleukin-3 receptor activity; interleukin-5 receptor activity; protein binding; protein-tyrosine kinase activity; Ras guanyl-nucleotide exchange factor activity; receptor activity

Biological Process: cellular protein metabolic process; MAPKKK cascade; respiratory gaseous exchange

Disease: Surfactant Metabolism Dysfunction, Pulmonary, 5

NCBI Summary:The protein encoded by this gene is the common beta chain of the high affinity receptor for IL-3, IL-5 and CSF. Defects in this gene have been reported to be associated with protein alveolar proteinosis (PAP). [provided by RefSeq, Jul 2008]
UniProt Code:P32927
NCBI GenInfo Identifier:1345923
NCBI Gene ID:1439
NCBI Accession:P32927.2
UniProt Secondary Accession:P32927,Q5JZI1, Q6ICE0,
UniProt Related Accession:P32927
Molecular Weight:97,976 Da
NCBI Full Name:Cytokine receptor common subunit beta
NCBI Synonym Full Names:colony stimulating factor 2 receptor beta common subunit
NCBI Official Symbol:CSF2RB  
NCBI Official Synonym Symbols:CD131; IL3RB; IL5RB; SMDP5; CDw131  
NCBI Protein Information:cytokine receptor common subunit beta
UniProt Protein Name:Cytokine receptor common subunit beta
UniProt Synonym Protein Names:CDw131; GM-CSF/IL-3/IL-5 receptor common beta subunit; CD_antigen: CD131
Protein Family:Cytokine receptor
UniProt Gene Name:CSF2RB  
UniProt Entry Name:IL3RB_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-IL-3Rbeta 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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