Recombinant wnt5a

Manufactured by R&D Systems

Sourced in United States

Recombinant Wnt5a is a laboratory protein produced using recombinant DNA technology. Wnt5a is a secreted signaling protein that plays a role in various cellular processes.

Automatically generated - may contain errors

Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions) Recombinant wnt3a, by R&D Systems (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Recombinant klotho, by R&D Systems (1 mentions) Rosiglitazone, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Microcystin lr, by Merck Group (1 mentions) Birb 0796, by Cell Signaling Technology (1 mentions) Pd184352, by Bio-Techne (1 mentions) U0126, by Cell Signaling Technology (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Buthionine sulfoximine bso, by Merck Group (1 mentions) Stemspan media, by STEMCELL (1 mentions) Recombinant sfrp5, by R&D Systems (1 mentions) Sfrp5, by R&D Systems (1 mentions) Wnt2b, by R&D Systems (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) β glycerophosphate, by Merck Group (1 mentions)

Close spelling variants of this product

Wnt5a (52 citations) Recombinant human Wnt5a (10 citations) Recombinant Wnt5a protein (4 citations) Recombinant human Wnt5a protein (2 citations) Recombinant human Wnt5a (rhWnt5a) (2 citations)

22 protocols using recombinant wnt5a

Wnt5A and Klotho Signaling Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were treated with 100 ng/ml of recombinant Wnt5A (R&D Systems,
cat. no. 645WN010CF) for 16 h or 10ng/ml of recombinant klotho (R&D Systems,
cat. no. 5334-KL-025 ) for 48 h. KN93 (Cayman, cat. no. 13319) was used at a
final concentration of 10μM and Rosiglitazone (Sigma-Aldrich, cat. no.
R2408-50MG) at 10μM for 48 h unless otherwise stated. BRAF resistance
subclones were seeded overnight in the absence of PLX4720 and then treated as
indicated.

Behera R., Kaur A., Webster M.R., Kim S., Ndoye A., Kugel CH I.I.I., Alicea G.M., Wang J., Ghosh K., Cheng P., Lisanti S., Marchbank K., Dang V., Levesque M., Dummer R., Xu X., Herlyn M., Aplin A.E., Roesch A., Caino C., Altieri D.C, & Weeraratna A.T. (2017). Inhibition of age-related therapy resistance in melanoma by rosiglitazone-mediated induction of Klotho. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(12), 3181-3190.

+ Open protocol

+ Expand

Cell Culture and Transfection Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

HEK293T and HepG2 from ATCC, and Huh7 from Dr. Kezhong Zhang, Wayne State University, were cultured in DMEM (Corning, NY) with heat inactivated 10% FBS (GIBCO), 1% sodium pyruvate and 1% penicillin/streptomycin. Transfection of cells was carried out 16–24 h after plating with Lipofectamine 2000 reagent and harvested in about another 24 h for analysis. Recombinant WNT5A was purchased from R&D systems (645-WN-010).

Bhattacharya A., Wei J., Song W., Gao B., Tian C., Wu S.A., Wang J., Chen L., Fang D, & Qi L. (2022). SEL1L-HRD1 ER-associated degradation suppresses hepatocyte hyperproliferation and liver cancer. iScience, 25(10), 105183.

+ Open protocol

+ Expand

Immortalized mouse preadipocyte models

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

White preadipocytes from wild type mice (Wt) and knockout (KO) mice lacking p38α (p38α^−/−), p38β (p38β^−/−), p38γ (p38γ^−/−), p38δ (p38δ^−/−), MKK3 (MKK3^−/−) and MKK6 (MKK6^−/−) kinases were immortalized by infection with SV40TpBABE-neo virus as previously described (Matesanz et al., 2017 (link)). The validity of these cell culture model has been previously supported (Matesanz et al., 2017 (link), 2018 (link)). In any case, we have reconfirmed the KO of MKK3 and MKK6 by western blot and those of p38 by qPCR (Supplementary Figure S1).
Cells were grown in Dulbecco’s Modified Eagle Medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco), l-glutamine (2 mM, Gibco), streptomycin (100 μg/ml, Gibco) and penicillin (100 U/ml, Gibco) and incubated at 37°C under a 5% CO₂/95% air atmosphere. Confluent cells were trypsinized and seeded in tissue dishes at a density of 6 × 10⁵ cells/ml. After 8 h, the medium was aspirated and replaced with fresh medium without sera. After 4–5 h, the medium was replaced with fresh medium containing DMSO (control cells) or the indicated concentrations of BIRB 0796 (Cell Signaling), Box5 (EMD Millipore), JNK-IN-8 (Calbiochem), U0126 (Cell Signaling), PD184352 (Tocris), Microcystin L-R (Sigma) or recombinant Wnt5a (R&D Systems) and the incubation was continued for a further 8 h.

Díaz-Chamorro S., Garrido-Jiménez S., Barrera-López J.F., Mateos-Quirós C.M., Cumplido-Laso G., Lorenzo M.J., Román Á.C., Bernardo E., Sabio G., Carvajal-González J.M, & Centeno F. (2022). Title: p38δ Regulates IL6 Expression Modulating ERK Phosphorylation in Preadipocytes. Frontiers in Cell and Developmental Biology, 9, 708844.

+ Open protocol

+ Expand

Wnt5A-mediated Cell Cycle Regulation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were treated with 100 ng/ml of recombinant Wnt5A (R&D Systems, Minneapolis, MN) for 16 hours unless otherwise stated. For cell cycle analysis, cells were treated with rWnt5A for 16 hours, and 1 hour before irradiation additional rWnt5A was added to the media. Cells were irradiated with either 10 Gy (Nordian Gammacell 40 extractor, 1.22 Gy/min) for initial experiments or 2.8 Gy (J.L. Sheppard Irradiator, 2.83 Gy/min) for all remaining experiments.

Webster M.R., Xu M., Kinzler K.A., Kaur A., Appleton J., O’Connell M.P., Marchbank K., Valiga A., Dang V.M., Perego M., Zhang G., Slipicevic A., Keeney F., Lehrmann E., Wood W I.I.I., Becker K.G., Kossenkov A.V., Frederick D.T., Flaherty K.T., Xu X., Herlyn M., Murphy M.E, & Weeraratna A.T. (2014). Wnt5A promotes an adaptive, senescent-like stress response, while continuing to drive invasion in melanoma cells. Pigment cell & melanoma research, 28(2), 184-195.

+ Open protocol

+ Expand

Isolation and culture of mouse hematopoietic stem cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For all experiments, lineage⁻, Sca-1⁺, c-kit⁺ (LSK) and LSK, CD150⁺, CD48⁻ cells were isolated as previously described [9 (link)]. Sorted LSK cells were cultured in serum-free StemSpan media (Stem Cell Technologies, Vancouver, BC, Canada, http://www.stemcell.com) supplemented with 25 ng/mL stem cell factor (SCF) and fms-like tyrosine kinase-3 ligand (Flt3L). Sorted and LSK, CD150⁺, CD48⁻ cells were cultured in serumfree StemSpan media (supplemented with 25 ng/mL SCF and thrombopoietin (TPO). All cytokines were obtained from Peprotech (Rocky Hill, NJ, http://www.peprotech.com). Sorted cells were incubated overnight before reseeding at 5 × 10⁴ cells (LSK) or 1.5 × 10⁴ cells (LSK, CD150⁺, CD48⁻) per milliliter in fresh media. Recombinant Wnt5a (R&D Systems, Minneapolis, MN, http://www.abgent.com) and anti-Ryk antibody (clone RB1491, Abgent, San Diego, CA) were added at 250 ng/mL and 1 μg/mL, respectively. Media were changed at 48 hours. For specific experiments, cells were treated with 0.1 mM buthionine sulfoximine (BSO) (Sigma Aldrich).

Povinelli B.J, & Nemeth M.J. (2014). Wnt5a Regulates Hematopoietic Stem Cell Proliferation and Repopulation Through the Ryk Receptor. Stem cells (Dayton, Ohio), 32(1), 105-115.

+ Open protocol

+ Expand

Wnt5a and SFRP5 Regulation in FLS

Check if the same lab product or an alternative is used in the 5 most similar protocols

RA td-FLS were seeded in 6-well plates at a density of 12 × 10⁴ cells/well and maintained at 5%CO₂ for 24h. Cells were treated with recombinant Wnt5a (300 ng/ml) (R&D systems^®) and/or recombinant SFRP5 (R&D systems^®) for 4h and 24 h. For the combined Wnt5a/SFRP5 treatment, SFRP5 was added 10 min before Wnt5a stimulation. Td-FLS were collected and used for RNA and/or protein extraction.

Mahmoud D.E., Kaabachi W., Sassi N., Mokhtar A., Moalla M., Ammar L.B., Jemmali S., Rekik S., Tarhouni L., Kallel-Sellami M., Cheour E, & Laadhar L. (2021). SFRP5 Enhances Wnt5a Induced-Inflammation in Rheumatoid Arthritis Fibroblast-Like Synoviocytes. Frontiers in Immunology, 12, 663683.

+ Open protocol

+ Expand

Modulation of Osteogenic Differentiation by Wnt Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were seeded on 24-well-plates (25,000 cells/well) and maintained in growth medium for 24 h. Subsequently, the medium was changed to osteo/odontogenic medium. The osteo/odontogenic medium consisted of growth medium supplemented with 50 µg/mL ascorbic acid (cat. no. A-4034, Sigma-Aldrich, St. Louis, MO, USA), 250 nM dexamethasone (cat. no. D8893, Sigma-Aldrich), and 5 mM β-glycerophosphate (cat. no. G9422, Sigma-Aldrich). In some experiments, the osteo/odontogenic medium was supplemented with either 1, 10, 50, 100, or 200 ng/ml recombinant WNT5A (R&D Systems Inc.) or 500 ng/ml WNT2b (R&D Systems Inc.) to activate Wnt signaling. In contrast, pre-treatment with Wnt inhibitors, 0.5 µg/ml DKK1 (R&D Systems Inc.), and 25 µM IWP-2 (Tocris Bioscience), was used to inhibit Wnt signaling.

Kornsuthisopon C., Chansaenroj A., Manokawinchoke J., Tompkins K.A., Pirarat N, & Osathanon T. (2022). Non-canonical Wnt signaling participates in Jagged1-induced osteo/odontogenic differentiation in human dental pulp stem cells. Scientific Reports, 12, 7583.

+ Open protocol

+ Expand

Mammary Epithelial Cell Spheroid Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols

Some 300–10,000 viable single cells were plated on Ultra Low Attachment 6-well or 96-well plates (Corning Incorporated Life Sciences) and cultured in MEGMTM mammary epithelial cell growth medium (Lonza) with or without recombinant Wnt5a (R&D system) at 100 ng/mL for 1–3 wk. Spheroids with sizes greater than 100 µm were counted using an inverted microscope (Nikon). A more detailed description of the reagents, biochemistry assays, cellular analysis, and animal studies are provided in SI Appendix, Materials and Methods.

Zhang S., Zhang H., Ghia E.M., Huang J., Wu L., Zhang J., Lam S., Lei Y., He J., Cui B., Widhopf GF I.I., Yu J., Schwab R., Messer K., Jiang W., Parker B.A., Carson D.A, & Kipps T.J. (2019). Inhibition of chemotherapy resistant breast cancer stem cells by a ROR1 specific antibody. Proceedings of the National Academy of Sciences of the United States of America, 116(4), 1370-1377.

+ Open protocol

+ Expand

Wnt5a and FZD7 siRNA Transfection

Check if the same lab product or an alternative is used in the 5 most similar protocols

In all, 100 000 cells were seeded into the wells of a 6-well plate. Recombinant Wnt5a of 200 ng/ml (R&D Systems, Minneapolis, MN, USA) was added immediately after seeding together with the FZD7 siRNA transfection reagents as described above. After 48 h of transfection, phase-contrast images were taken under × 10 magnification (Carl Zeiss Microscopy, Gottingen, Germany) to observe the phenotypic changes.

Asad M., Wong M.K., Tan T.Z., Choolani M., Low J., Mori S., Virshup D., Thiery J.P, & Huang R.Y. (2014). FZD7 drives in vitro aggressiveness in Stem-A subtype of ovarian cancer via regulation of non-canonical Wnt/PCP pathway. Cell Death & Disease, 5(7), e1346-.

+ Open protocol

+ Expand

Evaluating Cell Invasion and Wnt5a Silencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell invasion was analyzed using the same protocol as for cell migration, but with the use of matrigel pre‐coated cell culture inserts (BD Bioscience) with 8‐μm pore‐size membranes. The invasive capability was analyzed in the presence or absence of recombinant IL‐6 (20 ng/ml), Box5 (100 μM), SB203580 (Sigma; 10 μM) and recombinant WNT5A (R&D Systems; 0.2 μg/ml). Following siRNA silencing of WNT5A in HTB63 cells (Supplementary Figure 2A) there was a reduced number of invaded cells and therefore these experiments were manually counted using an inverted light microscope (Nikon TMS) instead of analyzed as described for the migration method.

Linnskog R., Jönsson G., Axelsson L., Prasad C.P, & Andersson T. (2014). Interleukin‐6 drives melanoma cell motility through p38α‐MAPK‐dependent up‐regulation of WNT5A expression. Molecular Oncology, 8(8), 1365-1378.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!