/, CCL chemokines/Mouse MIP-1 alpha (CCL-3) Recombinant

Mouse MIP-1 alpha (CCL-3) Recombinant

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$70.00$2,700.00

SKU: RKP10855 Tags: , , , , , , ,

Description

Accession
P10855
Source
Optimized DNA sequence encoding Mouse CCL3/Macrophage Inflammatory protein-1 alpha mature chain was expressed in Escherichia Coli.
Molecular weight
Native Mouse MIP-1alpha/CCL3 is generated by the proteolytic removal of the signal peptide and propeptide. This molecule has a calculated molecular mass of approximately 8 kDa. Recombinant MIP-1alpha is a monomer protein consisting of 69 amino acid residue subunits,and migrates as an approximately 8 kDa protein under non-reducing andreducing conditions in SDS-PAGE.
Purity
>95%, as determined by SDS-PAGE and HPLC
Biological Activity
Determined by its ability to chemoattract murine balb/c splenocytes using a concentration range of.0-50.0 ng/ml.

Protein Sequence
MKVSTTALAV LLCTMTLCNQ VFSAPYGADT PTACCFSYSR KIPRQFIVDY FETSSLCSQP GVIFLTKRNR QICADSKETW VQEYITDLEL NA
Endotoxin
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than.1 ng/µg(1EU/µg).
Presentation
Recombinant MIP-1 alpha was lyophilized from a.2 μm filtered solution in.5% glycine,.5% sucrose,.01% Tween80, mM Glutamic acid, pH.5.
Reconstitution
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than.1 mg/mL. This solution can then be diluted into other buffers.
Storage
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at° -° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Interactor
E9M5R0
Biological Process
Biological Process
Molecular function

Methods

Ccr1 ligands are induced after Candida infection and are chemotactic for kidney neutrophils ex vivo.

  • *

Binding and in vitro activity of murine 18V4F hybridoma antibody.

  • Directly coated chemokines were used here for direct comparisons, however in other experiments CCL3 showed a significantly enhanced signal when biotinylated and coated on streptavidin plates.

Binding and in vitro activity of murine 18V4F hybridoma antibody.

  • Directly coated chemokines were used here for direct comparisons, however in other experiments CCL3 showed a significantly enhanced signal when biotinylated and coated on streptavidin plates.

Flow cytometric analyses

  • PBMC were cultured in the presence of adefovir, tenofovir, and PMEO-DAPym and incubated at 37°C in a humidified, 5% CO2-controlled atmosphere.
  • The expression of cellular activation markers was measured after 3 days of culture.
  • Briefly, after washing the cells with PBS containing 2% FCS, we incubated them with PerCP-conjugated anti-CD4 mAb (clone SK3) in combination with PE-conjugated anti-CD25, anti-CD69, or anti-HLA-DR mAbs for 30 min at room temperature.
  • For aspecific background staining, cells were stained in parallel with Simultest Control IgG γ1/γ2a and PerCP-conjugated mouse IgG1.
  • Finally, the cells were washed with PBS, fixed with 1% formaldehyde solution, and analyzed with a FACSCalibur ; data were acquired with CellQuest software and further analyzed with the FLOWJO software .
  • The expression of the chemokine receptors was measured after 24 h, and PBMC were incubated with PerCP-conjugated anti-CD4 mAb (clone SK3) in combination with APC-conjugated anti-CXCR4 mAb (clone 12G5) and PE-conjugated anti-CCR5 (clone 2D7) or…

In vivo neutrophil recruitment into the cremaster muscle

  • Male C57Bl/6 mice of similar age were injected with an intrascrotal injection of 200 µl with either saline (sterile NaCl) or COAM (0.2 mg) 3 or 24 h prior to experiments.
  • At the time for experiment, mice were anaesthetized by spontaneous inhalation of isoflurane gas via an isoflurane pump ( 400 , ’s , , ) through a breathing mask containing a mixture of air and oxygen (total oxygen 40%) and ∼2.4% isoflurane.
  • The animals were placed on a water-heated operating table to maintain body temperature at ∼37°C.
  • The depth of anesthesia was controlled by regularly monitoring peripheral reflexes.
  • The cremaster muscle was prepared as previously described 2) was analyzed.
  • The cremaster muscles were saved for further analyses.
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