siRNA / RNAi /miRNA transfection Human Cal 27 cells

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Get tips on using Human Decorin DuoSet ELISA to perform ELISA Human - Decorin

Products R&D Systems Human Decorin DuoSet ELISA

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

RNA RNA isolation / purification Cells primary hematopoietic stem cells

Stem cells have the unique ability to self-renew or differentiate themselves into various cell types in response to appropriate signals. These cells are especially important for tissue repair, regeneration, replacement, or in the case of hematopoietic stem cells (HSCs) to differentiate into various myeloid populations. Appropriate signals refer to the growth factor supplements or cytokines that mediate differentiation of various stem cells into the required differentiated form. For instance, HSCs can be differentiated into dendritic cells (with IL-4 and GM-CSF), macrophages (with m-CSF) and MDSCs (with IL-6 and GM-CSF). Human pluripotent stem cells (hPSCs) and induced pluripotent stem cells (iPSCs) can be first cultured in neural differentiation media (GSK3𝛃-i, TGF𝛃-i, AMPK-i, hLIF) to form neural rosettes, which can be differentiated into neural or glial progenitors (finally differentiated into oligodendrocytes). Neural progenitors can be finally differentiated into glutaminergic (dibytyryl cAMP, ascorbic acid) and dopaminergic (SHH, FGF-8, BDNF, GDNF, TGF-𝛃3) neurons. Thus, it is important to first identify the self-renewing cell line: its source and its final differentiation state, followed by the supplements and cytokines required for the differentiation, and final use. Timelines are another thing that is considered. For instance, it takes 7-10 days to form neural rosettes from iPSCs and 3 days to differentiate neural progenitors to neurons. Finally, the stability for stem cell culture media varies. It is advised to make fresh media every time when differentiating HSCs to myeloid populations, whereas neural differentiation media may remain stable for two weeks when stored in dark between 2-8C.

Cell culture media Stem cell Differentiation media Differentiation of Human iPSCs into Basal Forebrain cholinergic neurons (BFCN)

Get tips on using MammoCult™ Human Medium Kit to perform 3D Cell Culture Media Primary human breast tumors-Mammospheres

Products STEMCELL technologies MammoCult™ Human Medium Kit

Get tips on using IntestiCult™ Organoid Growth Medium (Human) to perform 3D Cell Culture Media Human gastric cancer organoids

Products STEMCELL technologies IntestiCult™ Organoid Growth Medium (Human)

Get tips on using IntestiCult™ Organoid Growth Medium (Human) to perform 3D Cell Culture Media Human pancreatic cancer organoids

Products STEMCELL technologies IntestiCult™ Organoid Growth Medium (Human)

Get tips on using Human FGF-10 Antibody to perform Immunohistochemistry Human - FGF-10

Products R&D Systems Human FGF-10 Antibody

Get tips on using Human Dkk-1 ELISA to perform ELISA Human - Dkk-1

Products Raybiotech Human Dkk-1 ELISA

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase

Cellular assays Reporter gene assay β-galactosidase substrates human knee bone tissue

Wound healing assay can be challenging due to inconsistencies and variations while making a wound on the confluent cell monolayer, consequently leads to wounds of varying sizes and widths. Moreover, this assay causes damage to the cells that are at the edge of the wound, which can prevent cell migration into the wound site and healing. The best solution is to use the standard wound healing assay kits using either combs or inserts to make a defined wound field or gap and prevent the well-to-well variation in these assays.

Cellular assays Wound healing assay cell type human Caco-2

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