Cultrex RGF Basement Membrane Extract Summary
Gelling Assay - BME gels in less than 30 minutes at 37°C, and maintains the gelled form in culture medium for a minimum of 14 days at 37°C.
Murine Engelbreth-Holm-Swarm (EHS) tumor.
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||12 - 18 mg/ml.
|Shipping||The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Product is stable for a minimum of 3 months from date of shipment when stored at -20 °C in a manual defrost freezer. For optimal stability, store at -80 °C. Avoid freeze-thaw cycles.
Storage Buffer: Dulbecco’s Modified Eagle’s medium without phenol red, with 10 ug/ml gentamicin sulfate.
Sterility Testing: PathClear® - Negative by PCR test for mycoplasma; 17 bacterial and virus strains typically included in mouse antibody production (MAP) testing, plus 13 additional murine infectious agents including LDEV, for a total of 31 organisms and viruses. No bacterial or fungal growth detected after incubation at 37 °C for 14 days following USP sterility testing guidelines.
AlkalinePhosphatase (AP) staining was used to characterize the pluripotency of humaninduced pluripotent stem (iPS) cells cultured on Cultrex Reduced Growth FactorBME (Catalog # 3433-010-01). Images are shown in 4X and 10X magnification. Datacourtesy of GABAeron.
Background: Basement Membrane Extracts
Basement membranes are continuous sheets of specialized extracellular matrix that form an interface between endothelial, epithelial, muscle, or neuronal cells and their adjacent stroma. Basement membranes are degraded and regenerated during development and wound healing. They not only support cells and cell layers, but they also play an essential role in tissue organization that affects cell adhesion, migration, proliferation, and differentiation. Basement membranes provide major barriers to invasion by metastatic tumor cells. Cultrex® Basement Membrane Extract (BME) is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm (EHS) tumor. The extract gels at 37°C to form a reconstituted basement membrane. The major components of BME include laminin, collagen IV, entactin, and heparin sulfate proteoglycan. BME can be used in a multiple applications, under a variety of cell culture conditions, for maintaining growth or promoting differentiation of primary endothelial, epithelial, smooth muscle and stem cells. BME can also be utilized in cell attachment, neurite outgrowth, angiogenesis, in vitro cell invasion and in vivo tumorigenicity assays.
Citations for Cultrex RGF Basement Membrane Extract
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
Citations: Showing 1 - 10
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Hyaluronan-Binding Protein Involved in Hyaluronan Depolymerization Controls Endochondral Ossification through Hyaluronan Metabolism
Authors: M Shimoda, H Yoshida, S Mizuno, T Hirozane, K Horiuchi, Y Yoshino, H Hara, Y Kanai, S Inoue, M Ishijima, Y Okada
Am. J. Pathol, 2017;0(0):.
Syndecan-1 is a novel molecular marker for triple negative inflammatory breast cancer and modulates the cancer stem cell phenotype via the IL-6/STAT3, Notch and EGFR signaling pathways
Authors: SA Ibrahim, R Gadalla, EA El-Ghonaim, O Samir, HT Mohamed, H Hassan, B Greve, M El-Shinawi, MM Mohamed, M Götte
Mol. Cancer, 2017;16(1):57.
Transposon mutagenesis identifies genes that cooperate with mutant Pten in breast cancer progression
Proc. Natl. Acad. Sci. U.S.A., 2016;0(0):.
Wnt signaling promotes breast cancer by blocking ITCH-mediated degradation of the YAP/TAZ transcriptional coactivator WBP2
Cancer Res, 2016;0(0):.
The M33 G protein-coupled receptor encoded by murine cytomegalovirus is dispensable for hematogenous dissemination but is required for growth within the salivary gland.
Authors: Bittencourt F, Wu S, Bridges J, Miller W
J Virol, 2014;88(20):11811-24.
Protein kinase D1 mediates anchorage-dependent and -independent growth of tumor cells via the zinc finger transcription factor Snail1.
Authors: Eiseler T, Kohler C, Nimmagadda S, Jamali A, Funk N, Joodi G, Storz P, Seufferlein T
J Biol Chem, 2012;287(39):32367-80.
CLM94, a novel cyclic amide with anti-VEGFR-2 and antiangiogenic properties, is active against primary anaplastic thyroid cancer in vitro and in vivo.
Authors: Antonelli A, Bocci G, La Motta C, Ferrari S, Fallahi P, Ruffilli I, Di Domenicantonio A, Fioravanti A, Sartini S, Minuto M, Piaggi S, Corti A, Ali G, Di Desidero T, Berti P, Fontanini G, Danesi R, Da Settimo F, Miccoli P
J Clin Endocrinol Metab, 0;97(4):E528-36.
Vesicle-associated Membrane Protein 3 (VAMP3) Mediates Constitutive Trafficking of the Renal Co-transporter NKCC2 in Thick Ascending Limbs: ROLE IN RENAL FUNCTION AND BLOOD PRESSURE.
Authors: Caceres P, Mendez M, Haque M, Ortiz P
J Biol Chem, 0;291(42):22063-22073.
PI3K/mTOR dual inhibitor VS-5584 preferentially targets cancer stem cells.
Authors: Kolev V, Wright Q, Vidal C, Ring J, Shapiro I, Ricono J, Weaver D, Padval M, Pachter J, Xu Q
Cancer Res, 0;75(2):446-55.
Combination of an allosteric Akt Inhibitor MK-2206 with etoposide or rapamycin enhances the antitumor growth effect in neuroblastoma.
Authors: Li Z, Yan S, Attayan N, Ramalingam S, Thiele C
Clin Cancer Res, 0;18(13):3603-15.
Effects of ulipristal acetate on human embryo attachment and endometrial cell gene expression in an in vitro co-culture system.
Authors: Berger C, Boggavarapu N, Menezes J, Lalitkumar P, Gemzell-Danielsson K
Hum Reprod, 0;30(4):800-11.
Src as a Therapeutic Target in Biliary Tract Cancer.
Authors: Nam A, Kim J, Park J, Bang J, Jin M, Lee K, Kim T, Han S, Im S, Kim T, Oh D, Bang Y
Mol Cancer Ther, 0;15(7):1515-24.
Function of carbonic anhydrase IX in glioblastoma multiforme.
Authors: Proescholdt M, Merrill M, Stoerr E, Lohmeier A, Pohl F, Brawanski A
Neuro Oncol, 0;14(11):1357-66.
KLF6 Suppresses Metastasis of Clear Cell Renal Cell Carcinoma via Transcriptional Repression of E2F1.
Authors: Gao Y, Li H, Ma X, Fan Y, Ni D, Zhang Y, Huang Q, Liu K, Li X, Wang L, Gu L, Yao Y, Ai Q, Du Q, Song E, Zhang X
Cancer Res, 0;77(2):330-342.
Conditioned medium from human amniotic mesenchymal stromal cells limits infarct size and enhances angiogenesis.
Authors: Danieli P, Malpasso G, Ciuffreda M, Cervio E, Calvillo L, Copes F, Pisano F, Mura M, Kleijn L, de Boer R, Viarengo G, Rosti V, Spinillo A, Roccio M, Gnecchi M
Stem Cells Transl Med, 0;4(5):448-58.
Preclinical Antitumor Efficacy of BAY 1129980-a Novel Auristatin-Based Anti-C4.4A (LYPD3) Antibody-Drug Conjugate for the Treatment of Non-Small Cell Lung Cancer.
Authors: Willuda J, Linden L, Lerchen H, Kopitz C, Stelte-Ludwig B, Pena C, Lange C, Golfier S, Kneip C, Carrigan P, McLean K, Schuhmacher J, von Ahsen O, Muller J, Dittmer F, Beier R, El Sheikh S, Tebbe J, Leder G, Apeler H, Jautelat R, Ziegelbauer K, Kreft B
Mol Cancer Ther, 0;16(5):893-904.
Metformin Reduces Prostate Tumor Growth, in a Diet-Dependent Manner, by Modulating Multiple Signaling Pathways.
Authors: Sarmento-Cabral A, L-Lopez F, Gahete M, Castano J, Luque R
Mol Cancer Res, 0;15(7):862-874.
Harvest of superficial layers of fat with a microcannula and isolation of adipose tissue-derived stromal and vascular cells.
Authors: Trivisonno A, Di Rocco G, Cannistra C, Finocchi V, Torres Farr S, Monti M, Toietta G
Aesthet Surg J, 0;34(4):601-13.
Fluorescent Image-Guided Surgery with an Anti-Prostate Stem Cell Antigen (PSCA) Diabody Enables Targeted Resection of Mouse Prostate Cancer Xenografts in Real Time.
Authors: Sonn G, Behesnilian A, Jiang Z, Zettlitz K, Lepin E, Bentolila L, Knowles S, Lawrence D, Wu A, Reiter R
Clin Cancer Res, 0;22(6):1403-12.
Vesicle-associated membrane protein 2 (VAMP2) but Not VAMP3 mediates cAMP-stimulated trafficking of the renal Na+-K+-2Cl- co-transporter NKCC2 in thick ascending limbs.
Authors: Caceres P, Mendez M, Ortiz P
J Biol Chem, 0;289(34):23951-62.
PTEN inhibits PREX2-catalyzed activation of RAC1 to restrain tumor cell invasion.
Authors: Mense S, Barrows D, Hodakoski C, Steinbach N, Schoenfeld D, Su W, Hopkins B, Su T, Fine B, Hibshoosh H, Parsons R
Sci Signal, 0;8(370):ra32.
Loss of PPARgamma in endothelial cells leads to impaired angiogenesis.
Authors: Vattulainen-Collanus S, Akinrinade O, Li M, Koskenvuo M, Li C, Rao S, de Jesus Perez V, Yuan K, Sawada H, Koskenvuo J, Alvira C, Rabinovitch M, Alastalo T
J Cell Sci, 0;129(4):693-705.
The hippo pathway effector YAP regulates motility, invasion, and castration-resistant growth of prostate cancer cells.
Authors: Zhang L, Yang S, Chen X, Stauffer S, Yu F, Lele S, Fu K, Datta K, Palermo N, Chen Y, Dong J
Mol Cell Biol, 0;35(8):1350-62.
Grainyhead-like 2 downstream targets act to suppress epithelial-to-mesenchymal transition during neural tube closure.
Authors: Ray H, Niswander L
GPER mediates activation of HIF1alpha/VEGF signaling by estrogens.
Authors: De Francesco E, Pellegrino M, Santolla M, Lappano R, Ricchio E, Abonante S, Maggiolini M
Cancer Res, 0;74(15):4053-64.
Isogenic blood-brain barrier models based on patient-derived stem cells display inter-individual differences in cell maturation and functionality.
Authors: Patel R, Page S, Al-Ahmad A
J Neurochem, 0;142(1):74-88.
How does Cultrex® Basement Membrane Extract (BME) promote cell differentiation?
All epithelial and endothelial cells are in contact with a basement membrane matrix on at least one of their surfaces. By providing them with their natural matrix in vitro as a substrate for the cells that provides biological cues, the cells can assume a more physiological morphology (i.e. correct shape) and begin expression of cell-lineage specific proteins. Two-dimensional plastic surfaces, in combination with serum-containing media, cause cells to flatten, proliferate and de-differentiate.
What is the Tube Formation Assay?
The Tube Formation Assay is based on the ability of endothelial cells to form three-dimensional capillary-like tubular structures when cultured on a hydrogel of reconstituted basement membrane, such as Cultrex Basement Membrane Extract (BME).
What are the advantages of the Tube Formation Assay?
The Tube Formation Assay is the most widely used in vitro angiogenesis assay. The assay is rapid, inexpensive and quantifiable. It can be used to identify potentially angiogenic and anti-angiogenic factors, to determine endothelial cell phenotype, and to study pathways and mechanisms involved in angiogenesis. It can be performed in a high throughput mode to screen for a large number of compounds.
What cell types can be used in the Tube Formation Assay?
The Tube Formation Assay is specific for endothelial cells, either primary cells or immortalized cell lines. Only endothelial cells form capillary-like structures with a lumen inside. Other endothelial cell types form other structures.
What are the variables associated with the Tube Formation Assay?
The major variables associated with tube formation are composition of the Cultrex Basement Membrane Extract (BME) hydrogel, thickness of the hydrogel, cell density, composition of angiogenic factors in the assay medium, and assay period.
How do I reduce spontaneous formation of tubular structures on Cultrex BME in the absence of angiogenic factors?
Primary endothelial cells, such as Human Umbilical Vein Endothelial Cells (HUVECs) form capillary-like structures in the absence of added angiogenic factors less often than immortalized endothelial cells. Generally, reducing the number of cells per cm2 plated onto Cultrex BME will result in less background or spontaneous tube formation. Titrate the number of cells and find optimal conditions for your specific cell line. When endothelial cells fully form capillary structures in response to angiogenic activators, but not in their absence, you may proceed.
Which Cultrex Basement Membrane Extract (BME) should I use for the Tube Formation Assay?
Cultrex Reduced Growth Factor BME (RGF BME) is generally used for testing compounds that promote angiogenesis because formation of capillary-like structures (tubes) is significantly less compared to non-growth factor reduced varieties of Cultrex BME. The Cultrex In Vitro Angiogeneis Assay (Tube Formation) includes a qualified production lot of Cultrex RGF BME that exhibits reduced background tube formation in the absence of angiogenic factors.
Reviews for Cultrex RGF Basement Membrane Extract
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