Igfbp2

Manufactured by R&D Systems

Sourced in United States

IGFBP2 is a protein that binds to insulin-like growth factors (IGFs) and regulates their bioavailability and activity. It is involved in the regulation of cell growth, differentiation, and metabolism.

Automatically generated - may contain errors

Lab products found in correlation

Interleukin 6 (il 6), by R&D Systems (3 mentions) Heparin, by Merck Group (2 mentions) Leptin, by Thermo Fisher Scientific (2 mentions) Tgf β1, by R&D Systems (2 mentions) Ccl25, by R&D Systems (2 mentions) Gm csf, by R&D Systems (2 mentions) Cd40l, by R&D Systems (2 mentions) Mmp 3, by R&D Systems (2 mentions) Mmp 9, by R&D Systems (2 mentions) Synergy h1 hybrid reader, by Agilent Technologies (1 mentions) Igf 1, by R&D Systems (1 mentions) D luciferin k salt bioluminescent substrate, by PerkinElmer (1 mentions) Complete mini, by Roche (1 mentions) Dc protein assay kit, by Bio-Rad (1 mentions) Mini protean tgx precast gels 4 20, by Bio-Rad (1 mentions) Irdye secondary antibody, by LI COR (1 mentions) Advanblock fluor, by Advansta (1 mentions) Igf 2, by Merck Group (1 mentions) Trans blot turbo transfer system, by Bio-Rad (1 mentions) Gapdh, by HyTest (1 mentions)

13 protocols using igfbp2

Evaluating IGFBP2 Effects on Multiple Myeloma Cell Growth

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

The BM-dependent MM cell lines, M, D and I were established in our laboratory as previously described (Li et al, 2007 (link)). To study the effects of IGFBP2 on MM cell growth, the three luciferase-expressing BM-dependent MM cell lines were cultured in serum-free αMEM for 48 h in the presence of IGF1 (40 ng/ml) and/or IGFBP2 (5 μg/ml) (R&D Systems, Minneapolis, MN, USA). After 48 h, D-Luciferin K+ Salt Bioluminescent Substrate (PerkinElmer, American Fork, UT, USA) was added to each well (1.5 mg/ml), and the bioluminescence was measured at 175 nm with a Synergy H1 hybrid reader (BioTek, Winooski, VT, USA) as described (Li et al, 2007 (link)).

Mehdi S.J., Johnson S.K., Epstein J., Zangari M., Qu P., Hoering A., van Rhee F., Schinke C., Thanendrarajan S., Barlogie B., Davies F.E., Morgan G.J, & Yaccoby S. (2018). Mesenchymal stem cells gene signature in high-risk myeloma bone marrow linked to suppression of distinct IGFBP2-expressing small adipocytes. British journal of haematology, 184(4), 578-593.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Targets

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

Cell lysates were prepared in TXLB lysis buffer [50 mM Hepes, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS, 0.5 mM EDTA, 50 mM NaF, 10 mM Na₃VO₄, and protease inhibitor cocktail (cOmplete Mini, EDTA-free, Roche)], and volumes were adjusted according to protein concentration measurements (DC protein assay kit, Bio-Rad, 5000111). Separation was performed by gel electrophoresis (Mini-PROTEAN TGX Precast Gels 4-20%, Bio-Rad, 4561096), before transferring onto a nitrocellulose membrane (Trans-Blot Turbo Transfer System, Bio-Rad) and blocking with AdvanBlock-Fluor (Advansta, R-03729-E10). Primary antibodies in AdvanBlock-Fluor were incubated overnight at 4°C; IGFBP2 (R&D Systems, AF674), IGF-II (Millipore, 05-166-MI), GAPDH (Hytest, 5G4MAB6C5), and GFP (Thermo Fisher Scientific, A11122). Membranes were washed between primary and secondary antibody treatments with Tris-buffered saline with 0.1% Tween 20 (TBST). IRDye secondary antibodies (1:5000 diluted in TBST; LI-COR) were incubated for at least 1 hour at RT, before detection on an Odyssey fluorescence imager CLx (LI-COR). Densitometry analysis was performed in FIJI (NIH) by normalizing the signal to GAPDH, which was used as a loading control.

Conway J.R., Dinç D.D., Follain G., Paavolainen O., Kaivola J., Boström P., Hartiala P., Peuhu E, & Ivaska J. (2023). IGFBP2 secretion by mammary adipocytes limits breast cancer invasion. Science Advances, 9(28), eadg1840.

+ Open protocol

+ Expand

Quantification of IGF and IGFBP biomarkers

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

ELISA kits for Insulin-like growth factor I (IGF1, #ab211651, intra-assay coefficient of variation (CV) 2.3%) and Insulin-like growth factor-binding protein 3 (IGFBP3, #ab211652, intra-assay CV 3.1%) were purchased from Abcam (Cambridge, MA), and ELISA kit for Insulin-like growth factor-binding protein 2 (IGFBP2, #DGB200, intra-assay CV 5%) was from R&D Systems (Minneapolis, MN), and ELISA kit for Anti-mullerian hormone (AMH, #AL-124, intra-assay CV 3.7%) was purchased from AnshLabs (Webster, TX). The plasma samples from each time point of selected subjects were diluted with dilution factors of 1:80, 1:100, and 1:2000 for IGF1, IGFBP2, and IGFBP3 detection, respectively; and for AMH measurement, dilution factors of 1:10 and 1:500 were used for samples from females and males, respectively. All ELISA measurements were conducted according to manufacturer’s protocol. Duplicated wells were used to average the absorbance values and the concentration of target proteins in the plasma were calculated based on the calibration curve (R² = 0.9848, 0.9999, 0.9936 and 0.9971 for IGF1, IGFBP2, IGFBP3, and AMH, respectively.) and dilution factor after subtraction of background.

Liu C.W., Bramer L., Webb-Robertson B.J., Waugh K., Rewers M.J, & Zhang Q. (2016). Temporal Profiles of Plasma Proteome during Childhood Development. Journal of proteomics, 152, 321-328.

+ Open protocol

+ Expand

Expansion of Cryopreserved UCB Cells

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

Cryopreserved UCB mononuclear cells (4 × 10⁵cells/ml) were suspended in serum-free Stemspan® medium (Stemcell technologies, vancouver, BC, Canada) supplied with a standard cytokine combination of 100 ng/ml SCF, 50 ng/ml FL and 100 ng/ml TPO (all three cytokines purchased from Peprotech, Rocky Hill, NJ, USA) and with individually varied doses and combinations of IGFBP1, IGFBP2, IGF2 and ANGPTL3 (these four cytokines purchased from R&D Systems, Minnneapolis, MN, USA). The cells were inoculated on a passage 3 to 5 BM-derived mesenchymal stromal cell layer and cultured in 37°C incubator for 12 days. The expanded cells were harvested at the end of 12 days and the adherent cord blood cells were detached after 1 minute of incubation at room temperature with 0.25% trypsin–ethylenediamine tetraacetic acid.

Fan X., Gay F.P., Lim F.W., Ang J.M., Chu P.P., Bari S, & Hwang W.Y. (2014). Low-dose insulin-like growth factor binding proteins 1 and 2 and angiopoietin-like protein 3 coordinately stimulate ex vivo expansion of human umbilical cord blood hematopoietic stem cells as assayed in NOD/SCID gamma null mice. Stem Cell Research & Therapy, 5(3), 71.

+ Open protocol

+ Expand

In vitro HSC Expansion and Differentiation

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

HSCs were cultured under ‘expansion' conditions in U-bottom 96-well plates for 5 days. Cultures were maintained in Stemline II (Sigma) supplemented with 10 μg ml⁻¹ Heparin (Sigma), 100 ng ml⁻¹ SCF (R&D Systems), 2 ng ml⁻¹ Flt3 ligand (R&D), 20 ng ml⁻¹ TPO (R&D Systems), 10 ng ml⁻¹ FGF-1 (Invitrogen), 500 ng ml⁻¹ IGFBP2 (R&D Systems) and 100 ng ml⁻¹ AngL-3 (R&D Systems)31 (link). At the end of the culture period, cells were stained with TMRM and analysed or sorted by flow cytometry. To induce differentiation, HSCs were cultured in a basal medium (Stemline II containing 100 ng ml⁻¹ SCF and 2 ng ml⁻¹ Flt3 ligand) supplemented with 20 ng ml⁻¹ IL-3 (R&D Systems) and 100 ng ml⁻¹ IL-6 (R&D Systems). For some experiments, 5 μM carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) (Sigma) was added at the medium. FCCP stock solution was prepared by dissolving the powder in ethanol at 10 mM concentration.

Vannini N., Girotra M., Naveiras O., Nikitin G., Campos V., Giger S., Roch A., Auwerx J, & Lutolf M.P. (2016). Specification of haematopoietic stem cell fate via modulation of mitochondrial activity. Nature Communications, 7, 13125.

+ Open protocol

+ Expand

Purification and Transduction of Human Fetal CD34+ Hematopoietic Stem and Progenitor Cells

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

Human CD34⁺ HSCPs were purified from fetal livers as previously described^{17 (link)}. Briefly, tissue was dissected into 5mm³ pieces in digestion buffer containing DNase I and collagenase D, incubated at 37°C for 30 minutes and homogenized. Following top layering with Ficoll-Paque (GE Healthcare) interphase containing immune cells and CD34⁺ HSPCs was collected and washed with PBS. EasySep human CD34 positive selection kit (StemCell Technologies) was then used to purify CD34⁺ HSPCs. For viral transduction, lentivirus was produced by transient transfection of 293 cells with plasmids encoding VSVG, delta8.9 and pGL3-derived lentivirus plasmids encoding GFP or GFP and NPM1c. HSPCs were propagated in Stem Span media supplemented with Angiopoietin-like 5 (Angptl5, Abnova), human stem cell factor (SCF), human fibroblast growth factor (FGF, Invitrogen), insulin-like growth factor binding protein 2 (IGFBP2, R&D systems), heparin (Sigma) and thrombopoietin (R&D systems). Cytokines were reconstituted in PBS + 0.1% BSA. The use of human tissue in this study was approved by Institutional Review Board (IRB) at Massachusetts Institute of Technology.

Kaur M., Drake A.C., Hu G., Rudnick S., Chen Q., Phennicie R., Attar R., Nemeth J., Gaudet F, & Chen J. (2019). Induction and Therapeutic Targeting of Human NPM1c+ Myeloid Leukemia in the Presence of Autologous Immune System in Mice. Journal of immunology (Baltimore, Md. : 1950), 202(6), 1885-1894.

+ Open protocol

+ Expand

Adipose-Derived Stem Cell Differentiation

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

PV-ADSCs were seeded on gelatin-coated flasks and differentiated with medium (α-MEM with 10% FBS and 5 ng/mL TGF-β1 [R&D systems]) for indicated time. Leptin (Peprotech, 450-31) or IGFBP-2 (R&D Systems, 797-B2-025) at indicated concentrations were used to manipulate differentiation.

Gu W., Nowak W.N., Xie Y., Le Bras A., Hu Y., Deng J., Issa Bhaloo S., Lu Y., Yuan H., Fidanis E., Saxena A., Kanno T., Mason A.J., Dulak J., Cai J, & Xu Q. (2019). Single-Cell RNA-Sequencing and Metabolomics Analyses Reveal the Contribution of Perivascular Adipose Tissue Stem Cells to Vascular Remodeling. Arteriosclerosis, Thrombosis, and Vascular Biology, 39(10), 2049-2066.

+ Open protocol

+ Expand

In Vivo Signaling Pathway Modulation

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

EdU (Sigma, 1 mg) was intraperitoneally injected into mice twice at 12 h intervals at 1 day before sacrifice. DMSO, PP2 (Millipore, 1 mM), NBI31772 (TOCRIS, 0.3 mM) and WAY 316606 hydrochloride (TOCRIS, 2 mM) in 100 μl were intradermally injected into mice twice a day for 4 days. IWP-2 (Sigma, 133.33 μM), Sfrp1 (R&D Systems, 20 μg/ml) and Igfbp2 (R&D Systems, 5 μg/ml) in 100 μl were intradermally injected into mice once a day for 4 days. LY364947 (Wako, 1.84 mM) in 100 μl was intradermally injected into mice once a day for 6 days.

Ichijo R., Kobayashi H., Yoneda S., Iizuka Y., Kubo H., Matsumura S., Kitano S., Miyachi H., Honda T, & Toyoshima F. (2017). Tbx3-dependent amplifying stem cell progeny drives interfollicular epidermal expansion during pregnancy and regeneration. Nature Communications, 8, 508.

+ Open protocol

+ Expand

Immunocytochemistry of Cell Cultures

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

For cell culture experiments, cells were grown on sterile, gelatin-coated glass cover slips and were fixed at 24 h after the first passage with 4% paraformaldehyde for 20 min at room temperature. Cells were permeabilized and background blocked with 0.3% Triton X-100 + 10% NGS in PBS for 45 min at room temperature. Primary antibodies were diluted in 1% NGS + 0.3% Triton X-100 + 1% BSA in PBS and incubated on slides overnight at 4°C. The primary antibodies and dilutions used were as follows: LAMP-1 (1:1000, Cat# 14–1071-82, Invitrogen, CA, USA), MCP-1 (1:500, Cat# MA5–17040, Invitrogen), IGF-1 (1:250, Cat# PAJ-27207, Invitrogen), and IGFBP-2 (1:250, Cat# MAB797, R&D Systems, NE, USA). Slides were incubated with appropriate secondary antibodies at a 1:300 dilution (AlexaFluors, Thermo Fisher Scientific). Samples stained for actin were incubated with ActinGreen 488 Ready Probe (Cat# R37110, Invitrogen). Slides were mounted with mounting medium containing DAPI nuclear counterstain (Cat# S36938, Thermo Fisher Scientific). Slides with secondary but no primary antibodies were used as negative background controls.

Zbinden J.C., Mirhaidari G.J., Blum K.M., Musgrave A.J., Reinhardt J.W., Breuer C.K, & Barker J.C. (2021). The lysosomal trafficking regulator is necessary for normal wound healing. Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society, 30(1), 82-99.

+ Open protocol

+ Expand

Isolation and Culture of Bone Marrow Cells

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

To harvest bone marrow cells, femurs and tibiae from WT BL6 mice were flushed under sterile conditions. Cells were filtered through a 40 μm mesh and cultured in Teflon bags (PermaLife PL-30) for 6 d in DMEM supplemented with 5% FBS, or 5% serum derived from either LF/HF mice or lean/obese humans. Recombinant proteins (CCL25, CD40L, GM-CSF, IGFBP2, IL5, IL6, MMP3, MMP9, 100 ng ml⁻¹ for all; R&D Systems) or neutralization antibodies (IgG versus anti-GM-CSF, 0.1 μg ml⁻¹) were added as indicated. Treatment media were changed every other day. After 6 d, flow cytometry was performed using antibodies against murine CD45, CD11b and Gr1 (see Supplementary Table 3).

Quail D.F., Olson O.C., Bhardwaj P., Walsh L.A., Akkari L., Quick M.L., Chen I.C., Wendel N., Ben-Chetrit N., Walker J., Holt P.R., Dannenberg A.J, & Joyce J.A. (2017). Obesity alters the lung myeloid cell landscape to enhance breast cancer metastasis through IL5 and GM-CSF. Nature cell biology, 19(8), 974-987.

+ 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!