Log In Sign up
The largest database of trusted experimental protocols
> Chemicals & Drugs > Biologically Active Substance > Chemotactic Factors

Chemotactic Factors

Chemotactic Factors are signaling molecules that guide the movement and migration of cells, particularly immune cells, towards sites of infection or inflammation.
These factors play a crucial role in the body's innate and adaptive immune responses, facilitating the recruitment of leukocytes to areas of need.
Chemotactic Factors can be derived from a variety of sources, including microbes, damaged tissues, and activated immune cells.
Proper regulation and utilization of Chemotactic Factors is essential for maintaining homeostasis and mounting effective immune responses.
Researchers studying Chemotactic Factors can leverage PubCompare.ai's AI-driven platform to optimize their research protocols, easily locate the best published methods, and gain valuable insights to take thier studies to the next leval.

Most cited protocols related to «Chemotactic Factors»

1
Cell Chemotaxis Analysis Protocol
Video microscopy was performed over a time period of 24 hours when using FCS as chemoattractant or twelve hours when using FaDu cells as attractant source. The time-lapse interval was ten minutes. Cell tracking was performed using the ImageJ software (National Institutes of Health, Bethesda, USA) plugin "Manual Tracking" (Fabrice Cordelières, Institut Curie, Orsay, France). Each experiment was repeated three times, completely independent from each other. On average 40-53 cells were tracked per experiment. In order to further analyse and evaluate chemotactical processes, we developed the ImageJ plugin, which is available for free as a download from the NIH ImageJ homepage. The "Chemotaxis and Migration Tool" provides different types of graphs and statistical tests based on the experimental data. Exported ASCII (mandatory format) tables from "manual tracking" were directly imported into the software tool, and the cell trajectories were all extrapolated to (x, y) = 0 at time 0 h.
For quantification of chemotaxis and migration, several values were generated to evaluate directed cell migration: the centre of mass, the forward migration indices in directions parallel and perpendicular to the direction of the gradient, and the Rayleigh [23 ] test as well as the cell velocity and directness, where the latter values provide indication for how straight cells move.
Zengel P., Nguyen-Hoang A., Schildhammer C., Zantl R., Kahl V, & Horn E. (2011). μ-Slide Chemotaxis: A new chamber for long-term chemotaxis studies. BMC Cell Biology, 12, 21.
Full text: Click here
Publication 2011
Cells Chemotactic Factors Chemotaxis Microscopy, Video Migration, Cell
2
Chemotaxis Analysis using Insall Chamber
The inner well is filled with 12.5 µl control medium consisting of MV3 medium without FBS. The prepared cover slip is then inverted and carefully lowered onto the chamber resting one edge before lowering gently to ensure the central chamber remains bubble free. Excess medium is blotted from the edges of the cover slip with care to avoid moving the cover slip, which could result in shearing of the cells over the bridge. The cover slip is then sealed into place with 1∶1 Vaseline:Paraffin (Melting point 58–62°C, Sigma-Aldrich). The outer surface of the cover slip is then washed with distilled water and dried before wiping with lens cleaning tissue. The chamber is then inverted to fill the outer chemoattractant well with approximately 80 µl MV3 medium using a 20 µl microloading tip (Eppendorf). FBS is therefore used as the chemoattractant as in other studies [26] (link). Holding the Insall chamber at 45 degrees whilst filling allows any bubbles to escape through the drilled holes. The two holes are then sealed with a thin strip of water resistant tape to avoid evaporation. The chemotactic responses of cells overlying the bridge region can then be analysed after incubating at 37°C for one hour to allow the gradient to form.
Muinonen-Martin A.J., Veltman D.M., Kalna G, & Insall R.H. (2010). An Improved Chamber for Direct Visualisation of Chemotaxis. PLoS ONE, 5(12), e15309.
Full text: Click here
Publication 2010
Chemotactic Factors Lens, Crystalline Paraffin Tissues Vaseline
3
Microfluidic Chemotaxis and Haptotaxis Assays

Protocol full text hidden due to copyright restrictions

Open the protocol to access the free full text link

Publication 2012
Biological Assay Cardiac Arrest Cell-Matrix Junction Cell Culture Techniques Cell Migration Assays Cells Chemotactic Factors Chemotaxis Dextran Haptotaxis Medical Devices Microchip Analytical Devices Serum Syringes tetramethylrhodamine isothiocyanate
4
Fibronectin Coated Migration Assay
Following the sterile water rinse, devices that were destined for cell migration studies were filled with 0.2 mg/mL fibronectin (Millipore) in phosphate buffered saline (PBS) and incubated for at least 2 hours at 37 °C. Cells were trypsinized, counted and suspended in complete medium (DMEM containing 10% fetal bovine serum) at a density of 5 × 106 cells/mL. The migration devices were rinsed with complete medium and aliquots of 4 μL were added to the cell inlet ports, corresponding to 20,000 cells. The cells were then allowed to attach overnight before changing the medium.
Migration studies to assess the effect of the chemoattractant were carried out in the incubator. Twenty four hours after seeding NIH 3T3 mouse embryonic fibroblasts (MEF), images were taken of the migration devices. The medium was then changed in both reservoirs simultaneously: the medium on the side of the cells was replaced with complete medium, whereas the reservoir towards which the cells migrated was replaced with complete medium containing concentrations of PDGF varying from 0 to 200 ng/mL. Migration devices destined for live cell imaging were treated similarly: the medium was replaced 24 hours after seeding with phenol-red free medium containing 25 mM HEPES (Gibco). Immediately following medium change, the devices were covered with a coverslip seal the device and limit medium evaporation.
Davidson P.M., Sliz J., Isermann P., Denais C, & Lammerding J. (2015). Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments. Integrative biology : quantitative biosciences from nano to macro, 7(12), 1534-1546.
Publication 2015
Cells Chemotactic Factors Embryo Fetal Bovine Serum Fibroblasts Fibronectins HEPES Medical Devices Migration, Cell Mus NIH 3T3 Cells Phocidae Phosphates Platelet-Derived Growth Factor Saline Solution Sterility, Reproductive
5
Transepithelial Migration Assay for Cystic Fibrosis
PMN transepithelial migration and subsequent inflammation occur first in the small airways of CF patients, i.e., the bronchiolar region, which is lined with a microvilli-covered epithelial monolayer dominated by Club cells [3 (link)–5 (link)]. Therefore, to mimic PMN transmigration into the small airway lumen, we selected the H441 human Club cell line [20 (link)] to grow epithelial monolayers at air-liquid interface (ALI). To enable PMN loading in the lamina propria and transepithelial migration (Fig. 1A), we used Alvetex (Reinnervate) 200 μm-thick inert 3D scaffolds with >90% porosity (pore sizes of 36–40 μm, with interconnects of 12–14 μm). In brief, inserts were activated with 70% ethanol, coated overnight at 37ºC with rat-tail collagen I (3 mg/mL, Sigma) and seeded with H441 cells at 2.5×105 cells per 12-well insert. Cells were first grown in submerged cultures with DMEM/F12 supplemented with 10% heat-inactivated serum, penicillin, and streptomycin. After 2 days, cells were supplemented basally with serum-free DMEM/F12 with 10% Ultroser G (Pall Life Sciences) to establish ALI. Cultures were grown for 2 weeks at ALI and supplemented basally with fresh medium every 48 hours. For TM experiments, the ALI cultures were placed with the apical compartment exposed to RPMI, leukotriene B4 (LTB4, 100 nM), CXCL8 (100 ng/mL), formyl-methionine-leucine-phenylalanine (fMLF, 100 ng/mL), lipopolysaccharide (LPS, 500 ng/mL), or airway supernatant (ASN) from CF, HC, COPD, and LD subjects. TM experiments with 0.5–1 ×106 PMNs loaded onto the 200 μm-thick basal compartment of the Alvetex scaffold (situated upside), and allowed to migrate at 37°C at 5% CO2 through the collagen and epithelial layers into the apical compartment (situated downside, and bathed with either control medium with chemoattractant, or ASN). In some experiments, drugs were added to apical ASN and/or basal PMN suspensions. In other experiments, LPS-RS (competitive inhibitor of LPS binding to TLR4) was added to apical LPS or CF ASN. LPS and LPS-RS were purchased as ultrapure reagents from InvivoGen.
Forrest O.A., Ingersoll S.A., Preininger M.K., Laval J., Limoli D.H., Brown M.R., Lee F.E., Bedi B., Sadikot R.T., Goldberg J.B., Tangpricha V., Gaggar A, & Tirouvanziam R. (2018). PATHOLOGICAL CONDITIONING OF HUMAN NEUTROPHILS RECRUITED TO THE AIRWAY MILIEU IN CYSTIC FIBROSIS. Journal of leukocyte biology, 104(4), 665-675.
Publication 2018
Bronchioles Cell Lines Cells Chemotactic Factors Chronic Obstructive Airway Disease Collagen Collagen Type I CXCL8 protein, human Ethanol Homo sapiens Inflammation Lamina Propria Leucine Leukotriene B4 Microvilli N-Formylmethionine Patients Penicillins Pharmaceutical Preparations Phenylalanine Serum Streptomycin Tail Transepithelial Migration Ultroser G

Most recents protocols related to «Chemotactic Factors»

1
Transwell Invasion Assay for Cell Migration
Transwell membranes were coated with Matrigel (40 μg/mL, Corning) at 37 °C for 1 h. The cells treated as indicated were seeded into the precoated upper chambers at a density of 5 × 104 cells/mL in serum-free medium, and the lower chambers were filled with medium containing 20% FBS as the chemoattractant. After culturing for 24 h at 37 °C, the noninvasive cells on the upper surface were removed with a cotton swab, and the cells that migrated to the lower surface through the membrane were fixed with 4% paraformaldehyde for 10 min. The fixed cells were then stained with crystal violet (Beyotime Biotechnology) for 10 min, washed with tap water, and counted under an inverted microscope (Olympus).
Zhang J., Guo F., Li C., Wang Y., Wang J., Sun F., Zhou Y., Ma F., Zhang B, & Qian H. (2023). Loss of TTC17 promotes breast cancer metastasis through RAP1/CDC42 signaling and sensitizes it to rapamycin and paclitaxel. Cell & Bioscience, 13, 50.
Full text: Click here
Publication 2023
Cells Chemotactic Factors Gossypium matrigel Microscopy paraform Serum Tissue, Membrane Violet, Gentian
2
Transwell Migration and Invasion Assay
For Transwell migration assay, Ca9-22 (0.8×105), HSC-2 (0.8×105), HSC-4 (1.8×105), and HN22 (1.8×105) cells in serum-free medium were seeded in the upper chambers of 24 well plate with Collagen type I-coated PET membrane of 8.0-µm pore size (BD Biosciences). For invasion assay, culture inserts of 24-well plate were coated with Matrigel (BD Biosciences) in a 37°C incubator for 2 h. Ca9-22 (1.0×105), HSC-2 (1.0×105), HSC-4 (1.2×105), and HN22 (1.2×105) cells in serum-free medium were seeded in the upper chambers. Lower chambers were filled with the media containing 10% FBS as a chemoattractant. After incubation for 24 h, the non-migratory or non-invasive cells in upper chamber were removed with a cotton swab. Cells on the lower surface of the filter were fixed with 100% methanol for 2 min and stained with H&E solution. Images of migratory or invasive cells were captured under an inverted light microscope (CKX53; Olympus Corporation) and the number of migratory or invasive cells were counted in randomly selected areas in three different microscopic fields (magnification, ×100).
Kim J.H., Lee K.W., Ahn D.G., Oh K.Y, & Yoon H.J. (2023). Clinical significance of L1CAM expression and its biological role in the progression of oral squamous cell carcinoma. Oncology Reports, 49(4), 67.
Publication 2023
Biological Assay Cell Migration Assays Cells Chemotactic Factors Collagen Type I Gossypium Light Microscopy matrigel Methanol Microscopy Serum Tissue, Membrane
3
Cellular Migration and Invasion Assay
Cellular potential for migration and invasion of the 2D and 3D cells were assessed using 24-well Transwell chambers (8 mm; BD Biosciences). For migration assay, a total of 4×104 cells in 200-µl suspension were seeded into the Transwell chamber, while 1×105 cells were seeded in the Matrigel-coated upper chamber for the invasion assay. The lower compartment of the chamber was filled with 500 µl of chemoattractant [conditioned medium prepared from human lung fibroblasts (MRC-5)]. The cells were allowed to migrate or invade for 24 h, after which the non-migratory or non-invasive cells on the top of the filter were carefully removed with cotton swabs. Membrane containing migratory or invasive cells was immersed in 25% methanol for 15 min, developed with 500 µl of 0.5% crystal violet (MilliporeSigma) for 15 min, and acid-extracted with 100 µl of 0.1 N HCl in methanol. The absorbance was measured at 550 nm using a microplate reader.
Keeratichamroen S., Sornprachum T., Ngiwsara L., Ornnork N, & Svasti J. (2023). p‑STAT3 influences doxorubicin and etoposide resistance of A549 cells grown in an in vitro 3D culture model. Oncology Reports, 49(4), 71.
Publication 2023
Acids Biological Assay Cell Migration Assays Cells Chemotactic Factors Culture Media, Conditioned Fibroblasts Gossypium Homo sapiens Lung matrigel Methanol Migration, Cell Tissue, Membrane Violet, Gentian
4
Cytokine Profile in Alcohol-Exposed Brain
Brain samples from one hemisphere, except hippocampus, striatum and cerebellum, were dissected under microscope between 2 and 4 h after the last cycle of alcohol exposure, weighed and snap frozen at −80°C. Brain sample weighted around 100–130 mg and were transferred to MagNa Lyser tubes (F. Hoffmann-La Roche Ltd., Basel, Switzerland) containing 500 μL of tissue extraction reagent II (FNN0081, Thermo Fisher Scientific, Waltham, MA, USA) and 1x protease inhibitor cocktail (P8340, Merck KGaA, Darmstadt, Germany) and homogenized. The homogenization protocol consisted of two rounds of 25 s at 6000 rpm, with 90 s pause in ice. Samples were then spun at 16000 g at 4°C for 1 min to remove foam, transferred to Eppendorf tubes, and then centrifuged at 16000 g at 4°C for 20 min. Supernatant was aliquoted and frozen at −80°C. The total protein concentration was measured by Bradford assay (Merck KGaA, Darmstadt, Germany). The cytokine assay was performed using the ‘Mouse TH1/TH2 9-Plex Tissue Culture Kit’ [Meso Scale Diagnostics (MSD), Rockville, MA, USA], following manufacturer instructions. Brain samples were loaded 1:10 in phosphate buffer saline. The assay allowed detection of mouse interferon (IFN)-γ, interleukin (IL)-1β, IL-2, IL-4, IL-5, keratinocyte chemoattractant (KC)/human growth-regulated oncogene (GRO) also known as chemokine ligand 1 (CXCL1), IL-10, IL-12 (total), and tumor necrosis factor (TNF)-α with a dynamic range from 0.23 to 10000 pg/mL. Plates were read using MSD Sector Imager 6000 and data analyzed using MSD Workbench software. Final concentrations were adjusted to the brain sample weights and the protein concentrations.
Tonetto S., Weikop P., Brudek T, & Thomsen M. (2023). Behavioral and biochemical effects of alcohol withdrawal in female C3H/HeNRj and C57BL/6JRj mice. Frontiers in Behavioral Neuroscience, 17, 1143720.
Full text: Click here
Publication 2023
Biological Assay Brain Buffers Cerebellum Cerebral Hemispheres Chemokine Chemotactic Factors CXCL1 protein, human Cytokine Diagnosis Ethanol Freezing Homo sapiens IL1B protein, human Interferons Interferon Type II Interleukin-2 Interleukin-4 Interleukin-5 Interleukin-10 Interleukin-12 Keratinocyte Ligands Microscopy ML 23 Mus Oncogenes Phosphates Protease Inhibitors Proteins Saline Solution Seahorses Striatum, Corpus Tissues Tumor Necrosis Factor-alpha
5
Transwell Assay for Cell Migration
We seeded 5 × 104 GI-ME-N cells per insert (8 µm, ThinCert™ Greiner Bio-One) in 200 µl 1% FBS-supplemented media containing DMSO or VPF. In the lower chamber, 800–1000 µl of 10% FBS media were used as a chemoattractant. The cells were allowed to migrate for 22 h, after which a cotton swab was used to remove the non-migrating cells from the upper chamber. The inserts were then washed in PBS before fixation for 20 min, at RT in 4% PFA solution. Following repeated washing steps for PFA removal, the migrating cells were stained for 30 min with 0.1% Crystal violet solution (Chroma-Gesellschaft Schmid & Co.). The inserts were washed and left to air-dry before being imaged using a ZEISS AxioImagerM2 fluorescence microscope. For the quantitative measurement of migrating cells, the crystal violet dye was extracted by incubating the membranes in a 10% acetic acid solution for 20 min, on a rocking platform at RT. The extracted dye was then spectrophotometrically measured at 595 nm. The transwell migration assay was performed in technical duplicates and repeated at least three times.
Condurat A.L., Aminzadeh-Gohari S., Malnar M., Schider N., Opitz L., Thomas R., Menon V., Kofler B, & Pruszak J. (2023). Verteporfin-induced proteotoxicity impairs cell homeostasis and survival in neuroblastoma subtypes independent of YAP/TAZ expression. Scientific Reports, 13, 3760.
Full text: Click here
Publication 2023
Acetic Acid Cell Migration Assays Cells Chemotactic Factors Gossypium Microscopy, Fluorescence Sulfoxide, Dimethyl Tissue, Membrane Violet, Gentian

Top products related to «Chemotactic Factors»

1
Matrigel by BD
Sourced in United States, United Kingdom, Germany, China, Canada, Japan, Italy, France, Belgium, Australia, Uruguay, Switzerland, Israel, India, Spain, Denmark, Morocco, Austria, Brazil, Ireland, Netherlands, Montenegro, Poland
Matrigel is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, a tumor rich in extracellular matrix proteins. It is widely used as a substrate for the in vitro cultivation of cells, particularly those that require a more physiologically relevant microenvironment for growth and differentiation.
2
Transwell chamber by Corning
Sourced in United States, China, United Kingdom, Germany, Switzerland, Japan, Australia
Transwell chambers are a type of lab equipment used for cell culture and biological assays. They consist of a permeable membrane insert placed inside a well, allowing for the study of cell-cell interactions and the movement of molecules across a barrier. The core function of Transwell chambers is to provide a controlled environment for culturing cells and monitoring their behavior and permeability.
3
Matrigel by Corning
Sourced in United States, China, Germany, United Kingdom, Canada, Japan, France, Netherlands, Montenegro, Switzerland, Austria, Australia, Colombia, Spain, Morocco, India, Azerbaijan
Matrigel is a complex mixture of extracellular matrix proteins derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is widely used as a basement membrane matrix to support the growth, differentiation, and morphogenesis of various cell types in cell culture applications.
4
Crystal violet by Merck Group
Sourced in United States, Germany, China, Japan, United Kingdom, France, Sao Tome and Principe, Italy, Australia, Macao, Spain, Switzerland, Canada, Belgium, Poland, Brazil, Portugal, India, Denmark, Israel, Austria, Argentina, Sweden, Ireland, Hungary
Crystal violet is a synthetic dye commonly used in laboratory settings. It is a dark purple crystalline solid that is soluble in water and alcohol. Crystal violet has a variety of applications in the field of microbiology and histology, including as a staining agent for microscopy and in the gram staining technique.
5
Biocoat matrigel invasion chamber by BD
Sourced in United States, United Kingdom, Germany, Japan, China, Spain, Italy, Canada
The BD BioCoat Matrigel Invasion Chambers are a laboratory equipment used to assess the invasive potential of cells. The chambers are pre-coated with Matrigel basement membrane matrix, which acts as a barrier to mimic the extracellular matrix. Researchers can use these chambers to study the ability of cells to invade through this barrier.
6
Transwell insert by Corning
Sourced in United States, United Kingdom, Germany, China, Switzerland, Japan, France, Spain
Transwell inserts are a type of laboratory equipment used for cell culture applications. They consist of a porous membrane that separates two chambers, allowing for the study of interactions between cells or the passage of substances across the membrane. The core function of Transwell inserts is to facilitate the creation of a barrier between the two chambers, enabling researchers to analyze various cellular processes and transport mechanisms.
7
Transwell chamber by BD
Sourced in United States, China, United Kingdom, Germany, Brazil, Japan
Transwell chambers are a type of cell culture insert used for studying cell migration, invasion, and permeability. They consist of a porous membrane that separates the upper and lower chambers, allowing the passage of cells, molecules, or other materials between the two compartments.
8
Crystal violet by Beyotime
Sourced in China, United States, Japan, Germany
Crystal violet is a synthetic dye commonly used in laboratory settings. It is a dark purple crystalline solid that is soluble in water and alcohol. Crystal violet is often used as a staining agent in various biological and medical applications, including microscopy and cell identification.
9
Transwell plate by Corning
Sourced in United States, China, United Kingdom
Transwell plates are a type of cell culture insert system used for in vitro studies. They consist of a permeable membrane support that separates the upper and lower compartments of a well, allowing for the study of cell migration, transport, and co-culture experiments.

More about "Chemotactic Factors"

Chemotactic Factors, also known as chemoattractants or chemokines, are signaling molecules that guide the movement and migration of cells, particularly immune cells, towards sites of infection or inflammation.
These crucial regulators play a pivotal role in the body's innate and adaptive immune responses, facilitating the recruitment of leukocytes, such as neutrophils, macrophages, and lymphocytes, to areas in need.
Chemotactic Factors can be derived from a variety of sources, including microbes, damaged tissues, and activated immune cells.
These factors bind to specific receptors on the surface of target cells, triggering a cascade of signaling events that direct the cells towards the source of the chemoattractant gradient.
In laboratory research, Chemotactic Factor studies often involve the use of Matrigel, a complex extracellular matrix derived from mouse sarcoma cells, and Transwell chambers or BD BioCoat Matrigel Invasion Chambers.
These tools enable researchers to assess cell migration and invasion in response to various Chemotactic Factors.
The Crystal violet staining method is commonly employed to visualize and quantify the migrated or invaded cells.
Proper regulation and utilization of Chemotactic Factors is essential for maintaining homeostasis and mounting effective immune responses.
Researchers studying Chemotactic Factors can leverage PubCompare.ai's AI-driven platform to optimize their research protocols, easily locate the best published methods, and gain valuable insights to take thier studies to the next leval.