Antibodies against PRL-3 (sc-130355), Myeloid cell leukemia-1 (Mcl-1) (sc-819), IL-13Rα1 (sc-398831) and IL-13Rα2 (sc-134363) were bought from Santa Cruz Biotechnology (Dallas, Texas, USA). Antibodies against phosphorylated (p) STAT1 (#9167), total STAT1 (#14994), pSTAT6 (#9361), total STAT6 (#9362) and B cell lymphoma-extra large (Bcl-xL) (#2762) were from Cell Signaling Technology (Beverly, MA, USA). Antibodies against GAPDH and PRL-3 (ab52976) were from Abcam (Cambridge, United Kingdom). Recombinant human C-C motif chemokine ligand 19 (CCL19) and IL-13 were from Peprotech (Rocky Hill, NJ, USA). The plasmid pLKO (non-silencing control) and shRNA-pLKO against PRL-3 were a kind gift from dr. Jim Lambert (University of Colorado, Denver, CO, USA) [16 (link)]. Neutralizing antibodies against IL-6 (MAB2061) and IL-13 (MAB213) were from R&D Systems (Minneapolis, MN, USA). PRL-3 inhibitor I (#P0108) (5-[[5-Bromo-2-[(2-bromophenyl)methoxy]phenyl]methylene]-2-thioxo-4-thiazolidinone) was from Sigma-Aldrich (St. Louis, MO, USA), Analog 3 (#Z44389470) was from Enamine Store (Ukraine) and thienopyridone was a kind gift from Professor John S. Lazo (University of Virginia, Charlottesville, VA, USA).
Mab2061
Manufactured by R&D Systems
Sourced in United States
MAB2061 is a mouse monoclonal antibody that recognizes human Epidermal Growth Factor Receptor (EGFR). It is suitable for use in various immunological techniques such as Flow Cytometry, Immunoprecipitation, and Western Blotting.
Automatically generated - may contain errors
Lab products found in correlation
Mab208, by R&D Systems (3 mentions)
Sc 134363, by Santa Cruz Biotechnology (1 mentions)
Mab213, by R&D Systems (1 mentions)
Prl 3 inhibitor 1, by Merck Group (1 mentions)
Bcl xl, by Cell Signaling Technology (1 mentions)
Wnt3a 5036 wn, by R&D Systems (1 mentions)
Kg 501, by Merck Group (1 mentions)
247 ilb, by R&D Systems (1 mentions)
Mab201, by R&D Systems (1 mentions)
Lgk974, by Selleck Chemicals (1 mentions)
Sulfasalazine, by Merck Group (1 mentions)
Anakinra, by Amgen (1 mentions)
Ab14955, by Abcam (1 mentions)
Cyclin b1, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Cyclin a, by Merck Group (1 mentions)
Aurkb, by Abcam (1 mentions)
Hmga2, by Santa Cruz Biotechnology (1 mentions)
Lmna c, by Santa Cruz Biotechnology (1 mentions)
β actin, by Merck Group (1 mentions)
9 protocols using mab2061
1
Antibody Sources and Inhibitors for PRL-3 Signaling
2
Modulation of Wnt and Cytokine Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lines and patient derived samples were treated with the following prior to downstream assays: 50 ng/ml DKK1 (GF170, Milipore), 50 µg/ml Vantictumab (Oncomed), 100 nm Wnt3A (5036-WN, R&D systems), 100 µM LGK974 (S7143, Selleckchem), 10 ng/ml rIL15 (247-ILB, R&D systems), 10 ng/ml rIL1β (201-LB, R&D systems), 5 µg/ml IL1β neutralising antibody (MAB201, R&D systems), 5 µg/ml IL15 neutralising antibody (MAB-274, R&D systems), 5 µg/ml IL6 neutralising antibody (MAB2061, R&D systems), 5 µg/ml IL8 neutralising antibody (MAB208, R&D systems), 10 µg/ml Anakinra (Amgen, Cambridge, UK), 5 mM Sulfasalazine (Sigma), 10uM KG-501 (Sigma). Treatment times for individual assays are detailed below.
3
Western Blotting Antibodies Inventory
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies used for Western blotting were as follows: cyclin A (C4710; Sigma-Aldrich, St. Louis, MO), cyclin B1 (4135; Cell Signaling, Beverly, MA), cyclin D1 (2926; Cell Signaling), p16 (sc-759; Santa Cruz Biotechnology, Dallas, TX), p21 (sc-397; Santa Cruz Biotechnology), p53 (sc-126; Santa Cruz Biotechnology), HMGA2 (sc-30223; Santa Cruz Biotechnology), H3S10phos (ab14955; Abcam, Cambridge, UK), histone H3 (ab1791; Abcam), AURKB (ab2254; Abcam), β-actin (A5441; Sigma-Aldrich), Rb (9309; Cell Signaling), LMNA/C (sc-7292; Santa Cruz Biotechnology), LMNB1 (ab16048; Abcam), IL-6 (MAB2061; R&D Systems, Minneapolis, MN), IL-8 (MAB208, R&D Systems), and MMP3 (a kind gift from Gillian Murphy, University of Cambridge, Cambridge, United Kingdom; Allan et al., 1991 (link)).
4
Lymphatic Endothelial Cell-Tumor Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LEC are human dermal lymphatic microvascular endothelial cells purchased from Lonza (HMVEC-dLyAd, CC-2810, Verviers, Belgium). They were cultured in EGM2-MV medium (herein referred as complete medium) (CC-3202, Lonza) until confluence. Tumor cell lines used were derived from human skin carcinomas: (i) spontaneously immortalized human keratinocyte HaCaT cells, (ii) malignant keratinocyte HaCaT–II–4 cells, and (iii) metastatic keratinocyte HaCaT-A5-RT3 cells [32 (link),33 (link),35 (link)]. Tumor cells were maintained in Dulbecco's modified Eagle's medium (DMEM, 10938-025) complemented with 10% fetal bovine serum (10270-106), 1% glutamine (25030-123) and 1% penicillin–streptomycin (15140-122) (all from Thermofisher, MA). Serum-starved tumor cells were treated with recombinant human IL6 (I1395, Sigma, Belgium) and a neutralizing antibody against IL6 (2.5 μg/ml, MAB 2061, R&D Systems, MN). LEC transfections with siRNAs targeting STAT1 (ON-TARGETplus Human STAT1 (6772), L-003543-00-0005, Dharmacon, Co, USA) or control siRNAs (ON-TARGETplus Non-Targeting Pool, D-00180-10-05, Dharmacon) were performed using Interferin (409-50, Polyplus). All cells used were negative for mycoplasma contamination.
5
In vivo Angiogenesis Assay Using Gelatin Sponges
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gelatin sponges (Gelfoam, Pfizer, Ixelles, Belgium) were cut into small pieces (5 mm³) and incubated with cells as previously described [36 (link)]. Briefly, HaCaT-A5-RT3 or HaCaT–II–4 cells (2 × 106 cells per sponge) were added to gelatin sponge in the absence or presence of LEC (4 × 106 cell per sponge), in EBM2 serum-free medium. Human neutralizing anti-IL6 antibody (15 μg/ml, MAB2061, R&D Systems) or irrelevant mouse IgG2b antibody (15 μg/ml, MAB004, R&D Systems) (as negative control) was added to cell suspension. After 30 min of incubation, sponges were embedded in interstitial type I collagen gel (1.5 mg/ml, 41.254.02, Serva). Sponges were subcutaneously inserted into the ears of nude mice and left for 7 days [36 (link),37 (link)]. Anti-IL6 or IgG2b antibody was injected directly in the sponge each day with a Hamilton syringe. At the end of the experiment, sponges were harvested, incubated in 4% formol (11699408, VWR) for 4 h, dehydrated in ethanol and fixed in paraffin (X881.2, Leica).
6
Neutralizing Antibodies for Growth Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following were used as neutralizing antibodies: human FGF acidic neutralizing antibody (R&D Systems, AF232); human FGF basic neutralizing antibody (R&D Systems, AF232); human FGF-7 neutralizing antibody (R&D Systems, MAB251); human Il-6 neutralizing antibody (R&D Systems, MAB2061); human HGF neutralizing antibody (R&D Systems, AB-294-NA); human PDGF-AA neutralizing antibody (R&D Systems, MAB221); human PDGF-BB neutralizing antibody (R&D Systems, AB-220-NA); and human/mouse Wnt-3a neutralizing antibody (R&D Systems, MAB9025).
7
Inhibition of Inflammatory Cytokines in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDA-MB-231 cells or MCF-7 at the density of 5×105 cells/ml were treated by M protein at concentrations of 60 pmol/ml for 24 h, then the conditioned medium was collected. The conditioned medium was centrifuged at 1000 rpm in 5 minutes, following by another step at 2000 rpm in 20 minutes to remove the cell death and cell debris. In order to inhibit the inflammatory cytokines, neutralizing antibodies were added to conditioned medium, including human anti-TNFα antibody (MAB210, R&D Systems, Minneapolis, MN, USA), anti-IL6 antibody (MAB2061, R&D Systems) and anti-IL8 antibody (MAB208, R&D Systems) at concentrations of 10 μg/ml, 5 μg/ml and 5 μg/ml respectively.
8
Protein Expression Analysis via Immunoblotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence and immunoblotting, on SDS-PAGE gels were performed as reported previously (Hoare et al., 2016 (link)). The following antibodies were used in this study: anti-COX2 (Cell Signaling, 12282, 1:1000); anti-HRAS (Calbiochem, OP-23, 1:500); anti-IL1a (R&D systems, MAB200, 1:100); anti-IL6 (R&D systems, MAB2061, 1:250); anti-IL8 (R&D systems, MAB208, 1:500); anti-β-Actin (Sigma, A5441, 1:5000); anti-C/EBPβ (Santa-Cruz, sc-150, 1:500); anti-IkBα (Cell Signaling, 4814, 1:1000); anti-PGE2 (Abcam, ab2318, 1:100); anti-CyclinA2 (Sigma, C4710, 1:500).
9
Mast Cell Cytokine and Mediator Release
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human ROSA KITWT MCs 18, 19 were cultured exactly as previously described: they were cultured in Iscove's Modified Dulbecco's Medium (ref 21980-032, Gibco, Thermo Fisher Scientific, Waltham, Mass) including the following supplements (penicillin 1%, streptomycin 1%, sodium pyruvate 1%; Minimum Essential Medium Vitamin solution 1%; Minimum Essential Medium Non Essential Amino Acids Solution 2%; and L-Glutamine 1%, Insulin-Transferrin-Selenium 1%, Gibco; Bovine Serum Albumin 0.3% CHoKL 10%, Sigma-Aldrich, St Louis, Mo), in the presence or the absence of recombinant IL-33 (10 ng/mL) or of neutralizing anti-IL33 (10 mg/mL), IgE (2 mg/mL; ref 401152, Merck Millipore, Burlington, Mass). MCs were also cultured with quercetin (Q4951, Sigma-Aldrich), cromolyn (C0399, Sigma-Aldrich), and neutralizing IL-6 antibody (MAB2061, R&D Systems, Minneapolis, Minn). Levels of histamine (ELISA Kit, IBL International GmbH, Tecan Group, M€ annedorf, Switzerland) and total tryptase (a and b) (ImmunoCAP Tryptase, Thermo Fisher Scientific) were measured in culture supernatants 1 hour after stimulation. Levels of platelet-derived growth factor (PDGF) and of TGF-b (Milliplex, Merck Millipore) were measured 5 days after stimulation.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!