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Mouse CCL1 / TCA-3 ELISA Kit

SKU:
MOFI00019
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P10146
Sensitivity:
9.375pg/ml
Range:
15.625-1000pg/ml
ELISA Type:
Sandwich
Synonyms:
CCL1, I-309, P500, SCYA1, SCYA2, TCA-3, C-C motif chemokine 1, chemokine, C-C motif ligand 1, I-309, inflammatory cytokine I-309, P500, SCYA1Small-inducible cytokine A1, SISe, small inducible cytokine A1, I-309, homologous to Mouse Tca-3, T lymphocyt
Reactivity:
Mouse
Research Area:
Cell Biology
Signaling Molecule:
Chemokine
$779
Frequently bought together:

Description

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Mouse CCL1/TCA-3 ELISA Kit

The Mouse CCL1/TCA-3 ELISA Kit is specifically designed for precise and reliable detection of CCL1/TCA-3 levels in mouse serum, plasma, and cell culture supernatants. This ELISA kit is known for its high sensitivity and specificity, ensuring accurate and reproducible results for various research applications.CCL1/TCA-3 is an important chemokine involved in immune responses and inflammation, playing a key role in regulating the migration and activation of immune cells. Elevated CCL1/TCA-3 levels have been associated with various inflammatory conditions, making it a valuable biomarker for studying immune-related diseases and potential therapeutic interventions.

With its advanced technology and user-friendly protocol, the Mouse CCL1/TCA-3 ELISA Kit is an essential tool for researchers looking to explore the intricate mechanisms of immune regulation and inflammation in mouse models. Trust in this ELISA kit to provide precise and consistent results for your research needs.

Product Name:Mouse CCL1 / TCA-3 ELISA Kit
Product Code:MOFI00019
Size:96 Assays
Alias:CCL1, I-309, P500, SCYA1, SCYA2, TCA-3, C-C motif chemokine 1, chemokine, C-C motif ligand 1, I-309, inflammatory cytokine I-309, P500, SCYA1Small-inducible cytokine A1, SISe, small inducible cytokine A1, I-309, homologous to Mouse Tca-3, T lymphocyte-secreted protein I-309, TCA3
Detection Method:Sandwich ELISA
Application:This immunoassay kit allows for the in vitro quantitative determination of Mouse CCL1 concentrations in serum plasma and other biological fluids.
Sensitivity:9.375pg/ml
Range:15.625-1000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery: Matrices listed below were spiked with certain level of Mouse CCL1 and the recovery rates were calculated by comparing the measured value to the expected amount of Mouse CCL1 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)87-10594
EDTA plasma(n=5)85-10495
UFH plasma(n=5)87-10196
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Mouse CCL1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)87-100%86-104%88-101%
EDTA plasma(n=5)89-98%92-100%82-98%
UFH plasma(n=5)90-100%85-96%83-94%
Intra Assay:CV <8%
Inter Assay:CV <10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

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
UniprotP10146
UniProt Protein Function:CCL1: Cytokine that is chemotactic for monocytes but not for neutrophils. Binds to CCR8. Belongs to the intercrine beta (chemokine CC) family.
UniProt Protein Details:

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

Cellular Component: extracellular space; extracellular region

Molecular Function:protein binding; chemokine activity; cytokine activity

Biological Process: immune response; chemotaxis

UniProt Code:P10146
NCBI GenInfo Identifier:134512
NCBI Gene ID:20290
NCBI Accession:P10146.1
UniProt Secondary Accession:P10146,P14098, Q9QUR9, Q9QUS5,
UniProt Related Accession:P10146
Molecular Weight:9,836 Da
NCBI Full Name:C-C motif chemokine 1
NCBI Synonym Full Names:chemokine (C-C motif) ligand 1
NCBI Official Symbol:Ccl1  
NCBI Official Synonym Symbols:I-309; Scya1; Tca-3; BF534335  
NCBI Protein Information:C-C motif chemokine 1; P500; TCA3; CCR8 ligand; SIS-epsilon; T-cell activation protein 3; small inducible cytokine A1; small-inducible cytokine A1
UniProt Protein Name:C-C motif chemokine 1
UniProt Synonym Protein Names:P500; SIS-epsilon; Small-inducible cytokine A1; T-cell activation protein 3; TCA-3; TCA3
Protein Family:Cyclin
UniProt Gene Name:Ccl1  
UniProt Entry Name:CCL1_MOUSE

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

StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample (Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

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.

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