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Human C-C motif chemokine 28 (CCL28) ELISA Kit (HUEB0368)

SKU:
HUEB0368
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9NRJ3
Range:
78-5000 pg/mL
ELISA Type:
Sandwich
Synonyms:
MEC, CCL28, Mucosae Associated Epithelia Chemokine, VIC, chemokine, C-C motif ligand 28, member 28, MGC71902, Protein CCK1
Reactivity:
Human
€649
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Description

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Human C-C motif chemokine 28 (CCL28) ELISA Kit

The Human C-C Motif Chemokine 28 (CCL28) ELISA Kit is a reliable tool for the precise measurement of CCL28 levels in human serum, plasma, and cell culture supernatants. This kit provides exceptional sensitivity and specificity, ensuring consistent and dependable results for a variety of research studies.CCL28, also known as mucosae-associated epithelial chemokine (MEC), is a chemokine protein that plays a crucial role in immune response regulation and mucosal immunity.

It is involved in the recruitment and activation of immune cells, making it a key player in inflammatory processes and autoimmune disorders.By accurately measuring CCL28 levels, researchers can gain valuable insights into the mechanisms underlying immune responses and inflammatory conditions. This ELISA kit is an essential tool for studying the role of CCL28 in various diseases and developing potential therapeutic interventions.

Product Name:Human C-C motif chemokine 28 (CCL28) ELISA Kit
SKU:HUEB0368
Size:96T
Target:Human C-C motif chemokine 28 (CCL28)
Synonyms:Mucosae-associated epithelial chemokine, Protein CCK1, Small-inducible cytokine A28, MEC, SCYA28
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:78-5000pg/mL
Sensitivity:34pg/mL
Intra CV:4.8%
Inter CV:7.7%
Linearity:
Sample1:21:41:81:16
Serum(N=5)90-99%104-114%105-115%93-103%
EDTA Plasma(N=5)99-109%91-101%105-114%96-106%
Heparin Plasma(N=5)105-115%99-108%98-109%107-116%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum8080-85
Plasma8180-87
Function:Chemotactic activity for resting CD4, CD8 T-cells and eosinophils. Binds to CCR3 and CCR10 and induces calcium mobilization in a dose-dependent manner.
Uniprot:Q9NRJ3
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human C-C motif chemokine 28
Research Area:Immunology
Subcellular Location:Secreted
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:CCL28: Chemotactic activity for resting CD4, CD8 T-cells and eosinophils. Binds to CCR3 and CCR10 and induces calcium mobilization in a dose-dependent manner. Belongs to the intercrine beta (chemokine CC) family.
UniProt Protein Details:

Protein type:Chemokine; Secreted, signal peptide; Secreted; Motility/polarity/chemotaxis

Chromosomal Location of Human Ortholog: 5p12

Cellular Component: extracellular space; extracellular region

Molecular Function:chemokine activity

Biological Process: elevation of cytosolic calcium ion concentration; immune response; chemotaxis; positive regulation of cell-matrix adhesion; response to nutrient

NCBI Summary:This antimicrobial gene belongs to the subfamily of small cytokine CC genes. Cytokines are a family of secreted proteins involved in immunoregulatory and inflammatory processes. The CC cytokines are proteins characterized by two adjacent cysteines. The cytokine encoded by this gene displays chemotactic activity for resting CD4 or CD8 T cells and eosinophils. The product of this gene binds to chemokine receptors CCR3 and CCR10. This chemokine may play a role in the physiology of extracutaneous epithelial tissues, including diverse mucosal organs. Multiple transcript variants encoding two different isoforms have been found for this gene. [provided by RefSeq, Sep 2014]
UniProt Code:Q9NRJ3
NCBI GenInfo Identifier:12230650
NCBI Gene ID:56477
NCBI Accession:Q9NRJ3.1
UniProt Secondary Accession:Q9NRJ3,D7RIE7,
UniProt Related Accession:Q9NRJ3
Molecular Weight:12,375 Da
NCBI Full Name:C-C motif chemokine 28
NCBI Synonym Full Names:chemokine (C-C motif) ligand 28
NCBI Official Symbol:CCL28
NCBI Official Synonym Symbols:MEC; CCK1; SCYA28
NCBI Protein Information:C-C motif chemokine 28; CC chemokine CCL28; small inducible cytokine A28; small-inducible cytokine A28; mucosae-associated epithelial chemokine; chemokine (C-C motif) ligand 28 splice variant chi; small inducible cytokine subfamily A (Cys-Cys), member 28
UniProt Protein Name:C-C motif chemokine 28
UniProt Synonym Protein Names:Mucosae-associated epithelial chemokine; MEC; Protein CCK1; Small-inducible cytokine A28
Protein Family:Meckelin
UniProt Gene Name:CCL28
UniProt Entry Name:CCL28_HUMAN
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir

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

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.
Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.
ELISA
Human CCL28 / MCP-3 ELISA Kit

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