Nori Sheep CCL3 ELISA Kit

$461.00$832.00

DataSheet   

This ELISA kit is for quantification of CCL3 in sheep. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for CCL3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CCL3 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for CCL3 is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of CCL3 bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for CCL3: Chemokine (C-C motif) ligand 3 (CCL3), macrophage inflammatory protein 1-alpha (MIP-1-alpha)

This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.

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Description

Nori Sheep CCL3 ELISA Kit Summary

Alternative names for CCL3: Chemokine (C-C motif) ligand 3 (CCL3), macrophage inflammatory protein 1-alpha (MIP-1-alpha)

Alternative names for sheep: ovine, lamb, goat

Assay Type Solid Phase Sandwich ELISA
Format 96-well Microplate or 96-Well Strip Microplate
Method of Detection Colorimetric
Number of Targets Detected 1
Target Antigen Accession Number na
Assay Length 3 hours
Quantitative/Semiquantitative Quantitative
Sample Type Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL,
Recommended Sample Dilution (Plasma/Serum) No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ
Sensitivity 3 pg/mL
Detection Range 15.63-1000 pg/mL
Specificity Sheep CCL3
Cross-Reactivity < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested.
Interference No significant interference observed with available related molecules
Storage/Stability 4 ºC for up to 6 months
Usage For Laboratory Research Use Only. Not for diagnostic or therapeutic use.
Additional Notes The kit allows for use in multiple experiments.

 

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background:

Chemokine (C-C motif) ligand 3 (CCL3) also known as macrophage inflammatory protein 1-alpha (MIP-1-alpha) is a protein that in humans is encoded by the CCL3 gene. CCL3 is a cytokine belonging to the CC chemokine family that is involved in the acute inflammatory state in the recruitment and activation of polymorphonuclear leukocytes[1] through binding to the receptors CCR1, CCR4 and CCR5.[1] Sherry et al. (1988) demonstrated 2 protein components of MIP1, called by them alpha (CCL3, this protein) and beta (CCL4).[2] CCL3 produces a monophasic fever of rapid onset whose magnitude is equal to or greater than that of fevers produced with either recombinant human tumor necrosis factor or recombinant human interleukin-1. However, in contrast to these two endogenous pyrogens, the fever induced by MIP-1 is not inhibited by the cyclooxygenase inhibitor ibuprofen and CCL3 may participate in the febrile response that is not mediated through prostaglandin synthesis and clinically cannot be ablated by cyclooxygenase.[3] CCL3 has been shown to interact with CCL4.[4] Attracts macrophages, monocytes and neutrophils.

References

  1. Wolpe SD, et al. (1988). The Journal of Experimental Medicine 167 (2): 570–81.
  2. Sherry B, et al. (1988). The Journal of Experimental Medicine 168 (6): 2251–9.
  3. Davatelis G, et al. (1989). Science (New York, N.Y.) 243 (4894 Pt 1): 1066–8.
  4. Guan E, et al. (2001). The Journal of Biological Chemistry 276 (15): 12404–9.

 

 

 

Product Citation

40.        

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