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产品信息 >> 试剂盒相关 >> ELISA 试剂盒;
Human CCL3/MIP-1 alpha

Human beta-NGF DuoSet ELISA Summary

Assay   Type

Solid   Phase Sandwich ELISA

Format

96-well   strip plate

Assay   Length

3.5   hours or 4.5 hours

Sample   Type & Volume Required Per Well

Cell   Culture Supernates (10 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin   Plasma (200 uL), Citrate Plasma (200 uL)

Sensitivity

10 pg/mL

Assay   Range

31.2 -   1,000 pg/mL (Cell Culture Supernates), 46.9 - 1,500 pg/mL (Serum, EDTA   Plasma, Heparin Plasma, Citrate Plasma)

Specificity

Natural   and recombinant human MIP-1 alpha

 

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.

Product Summary

The Quantikine Human MIP-1 alpha Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure MIP-1 alpha levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human MIP-1 alpha and antibodies raised against the recombinant protein. Results obtained for naturally occurring human MIP-1 alpha showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of natural human MIP-1 alpha.

Preparation and Storage

Shipping

The   product is shipped at ambient temperature. Upon receipt, store it immediately   at the temperature recommended below.

Storage

Store   the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: CCL3/MIP-1 alpha

MIP-1 alpha (macrophage inflammatory protein 1 alpha) is a member of the CC or beta chemokine subfamily that was originally purified from the conditioned media of an LPS-stimulated murine macrophage cell line. MIP-1 alpha acts as a chemoattractant to a variety of cells including monocytes, T cells, B cells and eosinophils.

The two human MIP-1 alpha genes arise by duplication/mutation. They code for MIP-1 alpha isoforms CCL3/LD78a and CCL3L1/LD78b, which share 94% amino acid sequence homology. Whereas the human CCL3/LD78a is a single-copy gene, the human CCL3L1/LD78b gene copy number varies within the population. Human CCL3L1/LD78b binds and signals through chemokine receptors CCR1, CCR5. When compared to CCL3/LD78a, CCL3L1/LD78b has higher binding affinity to CCR5, which also functions as a coreceptor for HIV-1 entry. The copy number of CCL3L1 is one of several genetic determinants of HIV-1 susceptibility.

Long   Name:

macrophage   inflammatory protein 1 alpha

Entrez   Gene IDs:

6348   (Human); 20302 (Mouse); 25542 (Rat); 448787 (Canine); 102136134 (Cynomolgus   Monkey)

Alternate   Names:

C-C   motif chemokine 3; MIP1-(a); AI323804; CCL3; chemokine (C-C motif) ligand 3;   G0S19-1; LD78a; LD78alpha; MIP1 alpha; MIP-1 alpha; MIP1A; MIP-1alpha;   MIP1-alpha; PAT 464.1; SCYA3; SIS-beta

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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