Because ATRA could cause IFNα synthesis and secretion in NB4 cells by up - regulating IRF - 1 (23), we thus quantitated the IFNα in culture
supernatant of ATRA - treated NB4 cells.
The results revealed that
the supernatant of 72 - hour ATRA - treated NB4 cells was sufficient to induce the tyrosine phosphorylation of STAT2 and the endogenous RIG - G level in U3A cells, in comparison with the relative consistent level of total STAT2 (Fig. 3B).
RNA was also extracted from
supernatant of mosquitoes pools as described above and used for the ZIKV rRT - PCR assay.
The supernatant of apoptotic cells causes transcriptional activation of hypoxia - inducible factor — 1α in macrophages via sphingosine -1-phosphate and transforming growth factor - β.
Mcc was purified from the culture
supernatants of E. coli VCS257 cells harboring pJEM15 plasmid, as described earlier (Bieler et al., 2005).
(E and F) Using ELISA, IL - 2 and IFN - γ production were measured from culture
supernatants of (D) after 72 h of stimulation.
Cultured pericytes abundantly expressed calpain1 (Fig. 2d) and showed a significant induction of calpain1 activity after stimulation with
supernatants of Ang2 overexpressing HUVEC (Fig. 2e).
IFN - γ and IL - 2 production from
supernatants of individual wells was determined using an ELISA (BioLegend, San Diego, CA).
Furthermore, an increased amount of IL - 2 was present in
the supernatants of anti-CD3 — stimulated DKO and Cbl - b − / − — stimulated CD8 + T cells relative to DGKζ − / − CD8 + T cells, which was increased relative to WT CD8 + T cells (Fig. 3E).
Not exact matches
«It's an interesting project because backwash
supernatant disinfection is not widely used in Australia,» said Michael Bambridge, Managing Director
of CST Wastewater Solutions.
Hybridoma
supernatants were screened for anti-rGRF activity by use
of a pituitary culture assay system that can detect growth hormone - releasing factor in the femtomole range.
By securing a pellet (with neodymium magnets) on the sides
of the tube walls rather than at the bottom, it allows complete removal
of the
supernatant without touching the pellet.
With the current designs
of magnetic stands, these purifications can be problematic, often leaving carryover contaminants when the pellet is disturbed or when the
supernatant is not completely removed.
After achieving 80 — 90 % confluency, cell culture media were changed to maintenance media with 2 % FBS and were inoculated with 200 µL
of clinical sample or 100 µL
of passaged viral
supernatant.
To confirm the generation
of infectious virus, viral
supernatants were quantitated by an end - point dilution assay.
For each pool, 55 µL
of rabbit sera and 55 µL
of viral
supernatant at a concentration
of 100 TCID50 were mixed, incubated for 1 hour at 37 °C, and inoculated onto A549 cells in wells
of a 96 - well plate as described above.
To perform the virus neutralization assay, 55 µL
of viral
supernatant at a concentration
of 100 TCID50 and 55 µL
of serum (starting at a 1 ∶ 8 dilution) were mixed and incubated for 1 hour at 37 °C.
UNISI provides expertise in the measurement
of cell signalling proteins, such as cytokines, chemokines and inflammatory biomarkers in multiple samples (including serum, plasma and tissue culture
supernatants), using a multiplex suspension array system or a flow cytometric bead assay.
After the third wash, the
supernatant was discarded down to 100 µl and 650 µl
of Tris - HCL buffer was added followed by 250 µl
of fresh / frozen reduction buffer (100 mM Tris - Cl pH 7.4, 40 mM DTT).
Both technologies allow the simultaneous detection
of multiple factors in serum, plasma and tissue culture
supernatants.
This
supernatant was thawed on ice and 50 µl added to 400 µl HPLC grade water, 50 µl
of 50 % MPA and 200 µl
of HPLC grade heptane.
Phosphorylation
of supernatant protein factor enhances wits ability to stimulate microsomal squalene monooxygenase.
Angiopoietin - 2 protein levels
of cell culture
supernatants were measured by ELISA (R&D, #DANG20) according to the manufacturer's instructions.
Stimulation
of pericytes with
supernatants from either Ang2 overexpressing HUVEC or
supernatants from control HUVEC resulted in significantly increased pericyte migration following Ang2 stimulation (Fig. 2c, Supplementary Fig. 4c, d).
Gesicles can be collected from the
supernatant, yielding a concentrated stock
of your Cas9 - sgRNA gesicles.
In this study, we have developed a new sensitive and specific one step rRT - PCR for detection
of ZIKV in serum and cell culture
supernatants using a Taqman probe containing locked nucleotides, in the NS5 region
of ZIKV genome using sequences
of ZIKV strains circulating in Africa and Asia.
Briefly, the appropriate dilution
of tissue
supernatant was added to the cuvette containing 1.9 mL
of 50 mmol phosphate buffer (pH 7.0).
Briefly, 100 ml
of culture
supernatants were mixed with AVL - Carrier RNA buffer.
Spleen CD4 + T cells isolated from BMT recipients on day 21 were treated with vehicle solution or VPA for 24 h, the
supernatants were collected, and ELISA was performed to determine the level
of IFN - γ (C) and IL - 17A (D).
For the total input control, 20 %
of the total
supernatant was saved and frozen at − 80 °C.
Assessment
of hASC - CM composition found high expression
of various human growth factors (IL ‐ 6, 8, 12, eotaxin, IP10, MCP ‐ 1, VEGF, and TIMP ‐ 1) in the
supernatant following the co-culture
of hASCs with islet cells, while IP10, eotaxin, VEGF, and TIMP ‐ 1 became increased with time during islet co ‐ culture, suggesting the presence
of paracrine cross ‐ talk between islets and hASCs.
With current designs
of magnetic stands, these purifications can be problematic, often with carry over contaminants when the pellet is disturbed or when the
supernatant is not completely removed.
After blocking for 1 hour with 10 % normal goat serum (NGS), the slides were incubated with 200 μl primary antibody, either a polyclonal rabbit anti-Borrelia (Accurate chemicals, Westbury, NY) at 1:200 or the anti-OspA monoclonal, CB10, with a 1:30 dilution
of hybridoma
supernatant for 1 hour at room temperature (RT).
To characterize the isolate that replicated in HeLa cells, a 166 - bp RNA sequence containing the variable region C
of the envelope (Env) surface protein was PCR - amplified from infected HeLa cell tissue culture
supernatants.
1 ml
of antibody - containing hybridoma
supernatant is only $ 40, significantly cheaper than commercially available antibodies.
The cell culture
supernatant was collected and concentrated if necessary, and then 100 μL aliquots were used for detection with ELISA kit (PBL Biomedical Laboratories) following the protocol
of the manufacturer.
Cell
supernatants were discarded and cell pellets were lysed
of RBCs and then used for staining with antibodies for flow cytometry analysis.
Supernatants were stored at − 80 °C until the moment
of analysis.
Flow cytometric analysis
of BT474, MCF7 / HER2, or MCF7 target cells after incubation with
supernatant from HB1.F3.H2IgG, HB1.F3.Adeno - H2IgG, or HB1.F3.Lenti - H2IgG (C), followed by incubation with FITC - conjugated anti-human IgG (blue histograms).
Importantly, each
supernatant labeled MCF7 cells between one and two logs less than either
of the HER2 - overexpressing breast cancer cell lines.
UNISI will provide expertise in the measurement
of cell signalling proteins, such as cytokines, chemokines and inflammatory biomarkers in multiple samples (including serum, plasma and tissue culture
supernatants), using a multiplex suspension array system or a Cytometric bead assay.
0.1 mL
of supernatant was added to BHK - 21 and African green monkey kidney (Vero) cell cultures (ATCC, Manassas, VA) and monitored as above.
Modality
of access under this proposal: Users can provide human or animal serum, plasma or tissue samples, or cell pellets or
supernatants from cell models.
Single - cell suspension was obtained by filtering the
supernatant through a 40 - μm cell strainer, and cell suspension was then gently loaded onto a layer
of Histopaque - 1077 gradient (1 × 106 — 3 × 106 cells / mL HistoPaque in a total
of 3 - mL volume) and then centrifuged at 400 × g for 30 min at room temperature.
Afterwards, the
supernatant was removed by a short spin
of the plate bottom up at no more than 9 × g. To wash the pellet, 50 µl
of 70 % ethanol was pipetted into each well and again the plate was spun bottom up at no more than 9 × g for 5 — 10 sec.
The
supernatant was mixed with 300 µl
of Ni - NTA agarose (Qiagen), and the proteins were allowed to bind for 3 hours, followed by washing with 20 ml
of resuspension buffer, and eluted with 500 µl
of elution buffer (20 mM NaHPO4, 300 mM NaCl, 10 % glycerol, 150 mM Imidazole, pH 7.8).
After the
supernatant was filtered through 0.45 - μm membrane, it was added to target cells in the presence
of 8 μg / ml polybrene (Millipore, Billerica, MA).
In addition, we showed that, although the
supernatants from each
of the HCMV - infected cultures contained infectious virus, the viral copy number and the quantity and timing
of virus production varied among the various organ - specific MSC.
These changes were accompanied by severe EC damage, decreased E-cadherin RNA level, elevated IFN - gamma in splenocyte culture
supernatant, and production
of significant IgM antibody against intestinal microbiota.
The
supernatant, the medium in which the culture is grown, contains a multitude
of beneficial byproducts
of the growth process, including vitamins, enzymes, antioxidants, and immune stimulators.