Human Soluble CD4 Recombinant

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$160.00$1,950.00

SKU: RKP01730 Tags: , ,

Description

Accession
P01730
Source
Optimized DNA sequence encoding Human CD4 extracellular domain was expressed in HEK293 cells.
Molecular weight
Human soluble CD4is generated by the proteolytic removal of the extracellular domain and propeptide, the molecule has a calculated molecular mass of approximately 40 kDa. Recombinant CD4 is a monomer protein consisting of 371 amino acid residue subunits, and migrates due to glycosylation as an approximately 55kDa protein under reducing conditions in SDS-PAGE.
Purity
>90%, as determined by SDS-PAGE and HPLC
Biological Activity
The activity was tested by immobilized recombinant CD4 (1ug/ml) ability to support adhesion of HeLa cells (5x104 cells/well ) ,-80% will adhere aftermin atC.
Protein Sequence
MNRGVPFRHL LLVLQLALLP AATQGKKVVL GKKGDTVELT CTASQKKSIQ FHWKNSNQIK ILGNQGSFLT KGPSKLNDRA DSRRSLWDQG NFPLIIKNLK IEDSDTYICE VEDQKEEVQL LVFGLTANSD THLLQGQSLT LTLESPPGSS PSVQCRSPRG KNIQGGKTLS VSQLELQDSG TWTCTVLQNQ KKVEFKIDIV VLAFQKASSI VYKKEGEQVE FSFPLAFTVE KLTGSGELWW QAERASSSKS WITFDLKNKE VSVKRVTQDP KLQMGKKLPL HLTLPQALPQ YAGSGNLTLA LEAKTGKLHQ EVNLVVMRAT QLQKNLTCEV WGPTSPKLML SLKLENKEAK VSKREKAVWV LNPEAGMWQC LLSDSGQVLL ESNIKVLPTW STPVQP MALI VLGGVAGLLL FIGLGIFFCV RCRHRRRQAE RMSQIKRLLS EKKTCQCPHR FQKTCSPI
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 CD4 was lyophilized from a.2 μm filtered solution inPBS, pH.4.
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.
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Biological Process
Biological Process
Molecular function

Methods

Flow cytometry analysis

  • The following antibodies were used for cell surface molecules expression analysis: CD11c APC, CD80 FITC, CD83 FITC, CD40 PE, CD86 PE, MHC II (HLA-DR) FITC, MHC I (HLA-ABC) FITC, CD1a FITC, CD14 FITC, ILT3 PE, PD-L1 PE, TLR-2 PE, CCR5 PE, CCR7 PE, CXCR1 FITC, CXCR2 PE, CXCR4 PE, CD4 PE, CD4 PeCy7, IFNγ APC, all from eBioscience; CCR2 PerCP-Cy5.5 from BioLegend (San Diego, CA, USA) and CCR1 PE from R&D Systems (Minneapolis, MN, USA).
  • Cells were resuspended in PBS + 10% FBS and incubated with antibodies for 45 minutes, then washed twice with PBS + 3% FBS and fixed with IC Fixation Buffer (eBioscience).
  • GILZ PE antibody (eBioscience) was used for intracellular staining.
  • Cells were fixed, resuspended in Permeabilization Buffer (eBioscience) and incubated with the antibody for 20 minutes, then washed twice with Permeabilization Buffer and resuspended in flow cytometry buffer for data collection.
  • For IFNγ intracellular detection cells were stimulated with phorbol-12-myristate-13-acetate (PMA) (50 ng/ml) / ionomycin…

Flow cytometry analysis

  • The following antibodies were used for cell surface molecules expression analysis: CD11c APC, CD80 FITC, CD83 FITC, CD40 PE, CD86 PE, MHC II (HLA-DR) FITC, MHC I (HLA-ABC) FITC, CD1a FITC, CD14 FITC, ILT3 PE, PD-L1 PE, TLR-2 PE, CCR5 PE, CCR7 PE, CXCR1 FITC, CXCR2 PE, CXCR4 PE, CD4 PE, CD4 PeCy7, IFNγ APC, all from eBioscience; CCR2 PerCP-Cy5.5 from BioLegend (San Diego, CA, USA) and CCR1 PE from R&D Systems (Minneapolis, MN, USA).
  • Cells were resuspended in PBS + 10% FBS and incubated with antibodies for 45 minutes, then washed twice with PBS + 3% FBS and fixed with IC Fixation Buffer (eBioscience).
  • GILZ PE antibody (eBioscience) was used for intracellular staining.
  • Cells were fixed, resuspended in Permeabilization Buffer (eBioscience) and incubated with the antibody for 20 minutes, then washed twice with Permeabilization Buffer and resuspended in flow cytometry buffer for data collection.
  • For IFNγ intracellular detection cells were stimulated with phorbol-12-myristate-13-acetate (PMA) (50 ng/ml) / ionomycin…

mTECs protect and enhance Treg cell phenotype.

  • Purified CD4+ (a and b) or CD4+CD25+ T cells (c and d) were cultured alone or with mTECs for 3 days.
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