Mab645

Manufactured by R&D Systems

MAB645 is a monoclonal antibody product from R&D Systems. It is designed for research use.

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Lab products found in correlation

Anti wnt5a, by R&D Systems (2 mentions) Alexa fluor conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions) Ab166910, by Abcam (2 mentions) A5441, by Merck Group (2 mentions) Ab92544, by Abcam (2 mentions) Ab4047, by Abcam (2 mentions) Ab36969, by Abcam (2 mentions) Ab18180, by Abcam (2 mentions) Nbp1 04278, by Novus Biologicals (2 mentions) Ab8245, by Bio-Rad (2 mentions) Goat anti rat igg horseradish peroxidase conjugated haf005, by R&D Systems (2 mentions) Nb400 131, by Novus Biologicals (2 mentions) Anti lef1, by Santa Cruz Biotechnology (1 mentions) Anti tg2, by Cell Signaling Technology (1 mentions) Anti lamin a c sc 6215, by Santa Cruz Biotechnology (1 mentions) Anti phospho β catenin, by Cell Signaling Technology (1 mentions) Ab32572, by Abcam (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) Tgx stain free fastcast acrylamide kit, by Bio-Rad (1 mentions)

7 protocols using mab645

Investigating Wnt Signaling Pathway

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Anti LEF1 (sc374522, Santa Cruz); anti β−catenin (ab32572, AbCam); anti phospho β−catenin (9561, Cell Signaling); anti TG2 (3557, Cell signaling); anti TG2 (sc71632, Santa Cruz); anti Lamin A/C (sc-6215, Santa Cruz); anti Tubulin (T-4026, Sigma); anti Wnt5a (MAB645, R&D); anti Wnt10b (PA5-20530, Invitrogen).

Rossin F., Costa R., Bordi M., D’Eletto M., Occhigrossi L., Farrace M.G., Barlev N., Ciccosanti F., Muccioli S., Chieregato L., Szabo I., Fimia G.M., Piacentini M, & Leanza L. (2021). Transglutaminase Type 2 regulates the Wnt/β-catenin pathway in vertebrates. Cell Death & Disease, 12(3), 249.

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Apoptosis Analysis of CLL Cells

Cited 1 time

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Apoptotic analysis was performed as described [42 (link)]. CLL cells were cultured with or without NLCs or anti-Wnt5a antibody (MAB645; R&D Systems) (5 μg/mL) in RPMI culture-media, which were analyzed by Annexin V FITC Apoptosis Detection Kit I (BD Biosciences) and FACSCalibur Flow Cytometer (BD Biosciences). FlowJo (V10, FlowJo, OR, USA) was used for the analysis of apoptosis.

Guo Y., Pei H., Lu B., Zhang D., Zhao Y., Wu F., Sun H., Huang J., Li P., Yi C., Zhu C., Pan Y., Wu S., Chen C., Xu X, & Chen Y. (2022). Aberrantly expressed Wnt5a in nurse-like cells drives resistance to Venetoclax in chronic lymphocytic leukemia. Cell Death Discovery, 8, 82.

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Immunodetection of Wnt Signaling Proteins

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The immunocomplexes bound to the protein G sepharose beads obtained from CM-CITED2 and CM-Ctl as described above, were washed 5 times with PBS1× and immunopurified proteins were analyzed by western blot as previously described^{27 (link)}, with a rat monoclonal anti-WNT5A (MAB645, R&D Systems) or rabbit polyclonal anti-WNT11 (SC-50360, Santa Cruz) antibodies used at 1:1000 and 1:200 dilution, respectively. Loading was monitored by staining of PVDF membrane-bound proteins by ponceau s. Western blotting assays performed using 15 μg of protein extracts from zebrafish embryos harvested at 24 and 48 hpf after microinjection of either combined AUG and SPLICING (2.5 ng of each morpholino) or non-injected the control embryos (Control) prepared using conditions previously described^{27 (link)}. Proteins were separated by SDS-PAGE on TGX Stain-Free™ FastCast™ Acrylamide Kit (Bio-Rad). CITED2 protein was detected in zebrafish extracts using an anti-CITED2 rat monoclonal IgG_2A antibody (MAB5005, R&D System) used at 1:500 dilution as recommended by the manufacturer. Loading control was monitored by detection of total stained proteins transferred to the PVDF membrane.

Santos J.M., Mendes-Silva L., Afonso V., Martins G., Machado R.S., Lopes J.A., Cancela L., Futschik M.E., Sachinidis A., Gavaia P, & Bragança J. (2019). Exogenous WNT5A and WNT11 proteins rescue CITED2 dysfunction in mouse embryonic stem cells and zebrafish morphants. Cell Death & Disease, 10(8), 582.

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Immunoblotting and Immunofluorescence Antibodies

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In the experimental procedures described, the following antibodies were used: rabbit monoclonal anti-LRP1 (Ab92544; Abcam), mouse monoclonal anti-ABCA1 (Ab18180; Abcam), rabbit monoclonal anti-PSAP (ab166910; Abcam), mouse monoclonal anti-CTSD (NBP1-04278 Novus Biologicals), goat anti–SR-B1 (NB400-131; Novus Biologicals), rabbit anti-ABCG1 (ab36969; Abcam), rat anti–Wnt-5a (MAB645; R&D Systems), mouse monoclonal anti–β-actin (A5441, Sigma-Aldrich, 1:5,000), anti–α-tubulin (Ab4047, Abcam), anti-GAPDH (Ab8245), rabbit anti-GST (sc-459), goat anti-rabbit IgG-horseradish peroxidase conjugate (170–6515: Bio-Rad Laboratories,), goat anti-mouse IgG-horseradish peroxidase conjugate (170-6516; Bio-Rad Laboratories), and goat anti-Rat IgG-horseradish peroxidase conjugated (HAF005; R&D Systems). For immunofluorescence, Alexa Fluor–conjugated secondary antibodies and Alexa Fluor 488–conjugated and 568–conjugated antibodies were purchased from Jackson ImmunoResearch.

Oldoni F., van Capelleveen J.C., Dalila N., Wolters J.C., Heeren J., Sinke R.J., Hui D.Y., Dallinga-Thie G.M., Frikke-Schmidt R., Hovingh K.G., van de Sluis B., Tybjærg-Hansen A, & Kuivenhoven J.A. (2018). Naturally Occurring Variants in LRP1 (Low-Density Lipoprotein Receptor–Related Protein 1) Affect HDL (High-Density Lipoprotein) Metabolism Through ABCA1 (ATP-Binding Cassette A1) and SR-B1 (Scavenger Receptor Class B Type 1) in Humans. Arteriosclerosis, Thrombosis, and Vascular Biology, 38(7), 1440-1453.

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Antibody Characterization Protocol

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In the experimental procedures described the following antibodies were used: rabbit monoclonal anti-LRP1 (Ab92544; Abcam), mouse monoclonal anti-ABCA1 (Ab18180; Abcam), rabbit monoclonal anti-PSAP (ab166910; Abcam), mouse monoclonal anti-Cathepsin D (NBP1-04278 Novus Biologicals), goat anti-SR-B1 (NB400-131; Novus Biologicals), rabbit anti-ABCG1(ab36969; Abcam), rat anti-Wnt-5a (MAB645; R&D Systems), mouse monoclonal anti-β actin (A5441, Sigma-Aldrich, 1:5,000), anti-α tubulin (Ab4047, Abcam), anti-GAPDH (Ab8245), rabbit anti-GST (sc-459), goat anti-rabbit IgG-horseradish peroxidase (HRP) conjugate (170-6515: Bio-Rad Laboratories,), goat anti-mouse IgG-HRP conjugate (170-6516; Bio-Rad Laboratories), goat anti-Rat IgG Horseradish Peroxidase conjugated (HAF005; R&D Systems). For immunofluorescence, Alexa Fluor–conjugated secondary antibodies and Alexa Fluor 488 and 568–conjugated were purchased from Jackson ImmunoResearch.

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Mesomimetic Adhesion Assay for Ovarian Cancer Cells

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Human LP9 peritoneal mesothelial cells were grown on cover slips containing type I collagen gels (10 μg/mL) until confluency. RFP-tagged OVCAR3, OVCAR5, and OVCAR8 cells were serum starved overnight then treated with serum-free media, rWnt5a protein (0.4 μg/mL), LP9-CND, or LP9-Wnt5a^KO-CND for 24 hours. In some experiments, cells were pretreated with Wnt5a function-blocking antibody (MAB645, R&D Systems) or control IgG at 2 μg/mL for 24 hours prior to use. Cells (5 × 10⁴) from each treatment condition then were added atop of the confluent mesomimetic and allowed to adhere for 20 minutes and 1 hour, respectively. Mesomimetic cultures were washed with PBS; adherent cells were imaged with Echo Revolve fluorescent microscope and quantified using ImageJ software. The assay was done in three experimental replicates and repeated in three biological replicates for all conditions.

Asem M., Young A.M., Oyama C., De La Zerda A.C., Liu Y., Yang J., Hilliard T.S., Johnson J., Harper E.I., Guldner I., Zhang S., Page-Mayberry T., Kaliney W.J, & Stack M.S. (2020). Host Wnt5a Potentiates Microenvironmental Regulation of Ovarian Cancer Metastasis. Cancer research, 80(5), 1156-1170.

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Neuroinflammation Modulation Protocol

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Morphine sulfate was purchased from West-Ward (Eatontown, NJ). PLX5622-containing rodent diet (1 kg containing 1200 mg PLX5622) was purchased from Research Diets (New Brunswick, NJ). Ganciclovir sodium was purchased from Advanced ChemBlocks Inc. (Burlingame, CA). Interleukin 1 receptor antagonist (IL-1Ra) protein was purchased from R&D Systems Inc. (Minneapolis, MN). AC-YVAD-CMK, a selective inhibitor of caspase-1, JNK Inhibitor II (SP600125), NMDA receptor antagonist APV, selective astrocytic toxin L-α-aminoadipate (L-AA) as well as Gabapentin were purchased from Sigma-Aldrich (St. Louis, MO). Caspase-1 siRNA was purchased from Thermo Fisher Scientific (Waltham, MA). Antibodies used for immunoblotting were anti-IBa1 (1:1000; Wako, 016–20001); anti-GFAP (1:1000; Millipore, MAB360), anti-β-actin (1:1000; Santa Cruz Biotechnology, sc-1616-R), anti-IL-1β (1:500; Novus Biologicals, NB600–633), anti-Wnt5a (1:1000; R&D, MAB645), anti-caspase-1 (1:1000; Adiogen Corporation, AG-20B-0042). Antibodies used for immunohistochemistry were anti-GFAP (1:200; Millipore, AB5541) and anti-IL-1β (1:200; Novus Biologicals, NB600–633).

Edwards K.J., Metherell L.A., Yates M, & Saunders N.A. (2001). Detection of rpoB Mutations in Mycobacterium tuberculosis by Biprobe Analysis. Journal of Clinical Microbiology, 39(9), 3350-3352.

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