• The concentrations of TNF Alpha, CXCL12 and CCL2 were measured in cell supernatants by using ELISA kits in accordance with the manufacturer NA s protocols (TNF Alpha and CXCL12 from R&D systems; CCL2 from) on an ETIMax-3000 reader. Supernatants were collected 24 h after infection, spun at 10,000 rpm for 10 min to discard bacteria and cell debris, and stored at −80°C until assayed.
  • Primary crypts were cultured according to Sato et al. using reduced concentrations of murine recombinant-spondin1 (500ng/mL –& ) and varying concentrations of EGF. Organoid structures were imaged at day 6. Cell proliferation was measured by BrdU incorporation by incubation with 20 Mu M BrdU for 1 hour at 37°C before fixation.
  • Bio-Gel injection and selected by adherence, were tested for their production of pro-inflammatory (IL-6, IL-1 Beta and TNF-Alpha ) and anti-inflammatory (IL-10) cytokines in response to stimulation by LPS and IFN-Gamma, in the presence or not of graded concentrations of recombinant chemerin (from 10−12 to 10−6 M).
  • Isolated crypts were counted and embeded in matrigel ( 356231 growth factor reduced) that contains 1 Mu M Jagged (Ana-Spec) at 5–10 crypts/Mu l and cultured in a modified form of medium as described in. Briefly, MEM/F12 was supplemented by EGF 40 ng/ml, Noggin 200 ng/ml-spondin 500 ng/ml (&or ) , N-Acetyl-L-cysteine 1 Mu M and Y-27632 dihydrochloride monohydrate 20 ng/ml. cADPR, when indicated, was added to culture at 50 Mu M. 30–50 Mu l drops of matrigel with crypts were plated onto a flat bottom 48-well plate ( 3548) and allowed to solidify for 20 to 30 minutes in a 37°C C. 350 Mu l of crypt culture medium was then overlaid onto the matrigel, changed every other day, and maintained at 37°C in fully humidified chambers containing 6% CO2. Clonogenicity (colony-forming efficiency) was calculated by plating 50 to 400 crypts and assessing organoid formation 3 to 7 days after initiation…

Mouse CD34 protein (Recombinant) (STJP000636)

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STJP000636

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Host: HEK293
Note: STRICTLY FOR FURTHER RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Recombinant-Mouse CD34-protein was developed from hek293. For use in research applications.
Conjugation: Unconjugated
Formulation: 0.2 Mu m filtered PBS solution, pH7.2.
Dilution Range: Spin the vial and reconstite in distilled water to a concentration not less than 0.1 mg/mL. This can then be diluted into other buffers.
Storage Instruction: Can be stored in working aliquots at 2°C-8°C C for one month, or at-20°C C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles. NA
Endotoxin: Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg (1EU/µg). NA
Immunoreactivity: Measured by the ability to support the adhesion of the HUVEC cell line. Coated plates with immobilized recombinant mouse CD34 ( 0.8 Mu g/mL) are able to adhere 40% of cells ( 4 x 104 cells/well) after one hour at 37 °C NA
Gene Symbol: Cd34
Gene ID: 12490
Uniprot ID: CD34_MOUSE
Immunogen Region: ECD
Immunogen: Optimized DNA sequence encoding extracellular domain of mouse CD34 including a C-terminal 6His tag was expressed in HEK293 cells. NA
Tissue Specificity Expressed in the kidney where it is detected in the thin limb of Henle's loop (at protein level). Highly expressed in hematopoietic progenitor cell lines, brain and testis, and moderately in the thymus, spleen, and bone marrow, but not in adult liver.
Post Translational Modifications Highly glycosylated. Phosphorylated on serine residues by PKC.
Function Possible adhesion molecule with a role in early hematopoiesis by mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. Could act as a scaffold for the attachment of lineage specific glycans, allowing stem cells to bind to lectins expressed by stromal cells or other marrow components. Presents carbohydrate ligands to selectins.
Protein Name Hematopoietic Progenitor Cell Antigen Cd34
Cd Antigen Cd34
Database Links Reactome: R-MMU-198933
Cellular Localisation Membrane
Single-Pass Type I Membrane Protein
Alternative Protein Names Hematopoietic Progenitor Cell Antigen Cd34 protein
Cd Antigen Cd34 protein
Cd34 protein

Information sourced from Uniprot.org

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