Affi gel blue agarose beads

Manufactured by Bio-Rad

Sourced in United States

Affi-Gel blue agarose beads are a type of chromatography resin composed of agarose beads with a blue dye covalently attached. The core function of these beads is to facilitate the purification and separation of proteins and other biomolecules through affinity-based interactions.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions) Ascorbic acid, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Transferrin, by Merck Group (1 mentions) S nitroso n acetyl dl penicillamine snap, by Merck Group (1 mentions) Bgjb medium, by Thermo Fisher Scientific (1 mentions) Fgf18, by Thermo Fisher Scientific (1 mentions) Microslicer zero 1, by Ted Pella (1 mentions) L ascorbic acid, by Merck Group (1 mentions) Fgf1 antibody, by R&D Systems (1 mentions) Anisomycin, by Santa Cruz Biotechnology (1 mentions) Nsc23766, by Santa Cruz Biotechnology (1 mentions) Sp600125, by Merck Group (1 mentions)

Close spelling variants of this product

Affi-Gel Blue Gel 100–200 mesh crosslinked agarose beads Affi-Gel Blue Gel beads Affi-Gel Blue beads Affi-gel agarose Affi-Gel Blue Affi-Gel Blue Gel Affi-Gel beads Agarose beads Agarose gel Agarose

8 protocols using affi gel blue agarose beads

Incisor Proximal End Culture

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The proximal end of the incisor was dissected from one-month-old Gli1-LacZ mice and cultured with a Trowell culture system in vitro. Briefly, the proximal end of the incisor was cultured in BGJb media supplementary with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin (Invitrogen), and 0.1 mg/ml ascorbic acid (Sigma). For IAN implantation, the IAN was dissected out and surrounded by the proximal end. For neutralizing FGF1, we used Affi-Gel blue agarose beads (BioRad) and soaked them in FGF1 antibody (0.5 μg/ul, R&D system). Normal IgG control (0.5 μg/ul, R&D system) was used as control. For bead implantation, we used Affi-Gel blue agarose beads (BioRad) and soaked them in recombinant FGF1 protein (0.5 μg/μl, R&D system). BSA (0.5 μg/ul) was used as a control. The beads were put around the proximal end. Tissues were harvested after 3 days of culture and fixed in 4% paraformaldehyde.

Pei F., Ma L., Jing J., Feng J., Yuan Y., Guo T., Han X., Ho T.V., Lei J., He J., Zhang M., Chen J.F, & Chai Y. (2023). Sensory nerve niche regulates mesenchymal stem cell homeostasis via FGF/mTOR/autophagy axis. Nature Communications, 14, 344.

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Intradermal Injection of rhSCUBE3 Protein

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Intradermal injections of recombinant human SCUBE3 (rhSCUBE3) protein were performed as previously described (Plikus et al., 2008 (link)). Briefly, rhSCUBE3 (R&D Systems) was reconstituted in 0.1% BSA to a final concentration of 250 μg/ml. Affi-gel blue agarose beads (Bio-Rad) were washed three times in sterile 1x PBS, air dried, and resuspended with rhSCUBE3 or BSA. Beads were micro-implanted into dorsal skin using a 26G needle at P48. Beads were resupplied with recombinant proteins at 24, 48, and 72 hours using a glass microneedle injector. Skin was analyzed on day 14 after initial bead micro-implantation.

Liu Y., Guerrero-Juarez C.F., Xiao F., Shettigar N.U., Ramos R., Kuan C.H., Lin Y.C., de Jesus Martinez Lomeli L., Park J.M., Oh J.W., Liu R., Lin S.J., Tartaglia M., Yang R.B., Yu Z., Nie Q., Li J, & Plikus M.V. (2022). Hedgehog signaling reprograms hair follicle niche fibroblasts to a hyper-activated state. Developmental cell, 57(14), 1758-1775.e7.

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Kidney Capsule Transplantation of Tooth Germs

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Kidney capsule transplantation was performed as described previously (Oka et al., 2007 (link)). After genotyping, the tooth germs of the mandibular first molars were collected from P4.5 Ror2^fl/fl control or Osr2-Cre;Ror2^fl/fl mice. Each tooth germ was surgically transplanted under the kidney capsule of a host mouse. The explants were harvested after 21 days. For the rescue experiment, affi-Gel blue agarose beads (Bio-Rad) were soaked in BSA (100 μg/ml) or the Cdc42 activator (50 μg/ml; Cytoskeleton, Inc.) overnight at 4°C before use. After that, BSA and activator beads were transplanted into the kidney capsule with the tooth germ collected from Ror2^fl/fl control or Osr2-Cre;Ror2^fl/fl mouse mandibles at P4.5. The explants were collected after 2, 3 or 21 days.

Ma Y., Jing J., Feng J., Yuan Y., Wen Q., Han X., He J., Chen S., Ho T.V, & Chai Y. (2021). Ror2-mediated non-canonical Wnt signaling regulates Cdc42 and cell proliferation during tooth root development. Development (Cambridge, England), 148(2), dev196360.

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Palatal Development: Culture and Bead Assay

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Palatal shelf explant culture and bead implantation experiments were carried out using a previously described protocol with minor modification [28 (link)]. Briefly, Timed pregnant mice were sacrificed on post-coital day 13.0 (E13.0). The embryonic maxillary processes with the secondary palatal shelves were manually microdissected and cultured in BGJb medium supplementary with 10 U/ml penicillin/ streptomycin (Invitrogen), 50 mM transferrin (Sigma) and 150 μg/ml ascorbic acid (Sigma). For bead implantation, Affi-Gel blue agarose beads (BioRad) were soaked in recombinant Fgf18 proteins (1mg/ml, Peprotech), or BSA (1mg/ml) as control. Tissues were harvested after 24 hours of culture at 37°C at an atmosphere of 5% CO2 and 100% humidity and fixed in 4% paraformaldehyde for whole mount in situ hybridization experiments.

Xu J., Liu H., Lan Y., Aronow B.J., Kalinichenko V.V, & Jiang R. (2016). A Shh-Foxf-Fgf18-Shh Molecular Circuit Regulating Palate Development. PLoS Genetics, 12(1), e1005769.

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Xenopus Craniofacial Development Assays

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Peptides (Thermo Scientific) were designed according to predicted sequences including 9 amino acid (AA) Xenopus Bradykinin (xBdk) (SYKGLSPFR) and 8AA Des-Arg xBdk (SYKGLSPF), and diluted to 0.1 mg/ml or 0.2mg/ml. Affi-gel blue agarose beads (50–100 mesh, Bio-Rad) loaded with peptides were prepared according to Carmona-Fontaine, Thesis, 2011 . For rescues, beads resuspended in 0.1mg/ml peptide solution were implanted in the presumptive mouth region at stage 22 and scored at stage 40. For NC assays, beads resuspended in 0.2mg/ml peptide solution were implanted in side of head or presumptive mouth at stage 20–22. Embryos were fixed at tailbud (st. 26) for in situ hybridization analysis. For peptide-rescue assays, partial LOF morphants were employed to maximize viability.
NO donor, S-Nitroso-N-acetyl-DL-penicillamine (SNAP) (Sigma) was diluted to 100 mM in a 50% DMSO solution. For early rescues, 1nl of SNAP was coinjected with 17ng of morpholino into one-cell stage embryos. For late rescues (st. 20), 2–3nl of SNAP was injected into the presumptive mouth region. The nNOS inhibitor, TRIM (Sigma- T7313), was diluted to 1M concentration in DMSO, and applied to late neurula (st. 20) embryos. Embryos were collected at tailbud (st. 26) for sox9 in situ hybridization and at swimming tadpole (st. 40) for craniofacial morphology.

Jacox L., Sindelka R., Chen J., Rothman A., Dickinson A, & Sive H. (2014). The Extreme Anterior Domain Is an Essential Craniofacial Organizer Acting through Kinin-Kallikrein Signaling. Cell reports, 8(2), 596-609.

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Runx2 Conditional Knockout Molar Transplantation

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Gli1‐Cre^ERT2;Runx2^fl/fl and Runx2^fl/fl littermate control mice were injected with tamoxifen at PN3.5 and euthanized 2 days later. Whole mandibular first molars were carefully dissected and placed in PBS on ice. Host mice were anesthetized using isoflurane, then fur on the back was shaved and the kidney on the left side was exposed through a skin incision. The kidney capsule was opened using fine‐tip forceps. Two explants were transplanted under the kidney capsule of one host. Three weeks later, the explants were harvested for histological analysis. For the rescue experiment, Affi‐Gel blue agarose beads (Bio‐Rad Laboratories; 1537301) were washed in PBS and then incubated in recombinant mouse NOTUM protein (100 μg/mL; R&D Systems, Minneapolis, MN, USA; 9150‐NO) or bovine serum albumin (BSA) (100 μg/mL) for 1 hour at 37°C before transplantation. NOTUM beads or BSA beads were then applied to the explants from Gli1‐Cre^ERT2;Runx2^fl/fl mice and transplanted under the kidney capsule.

Wen Q., Jing J., Han X., Feng J., Yuan Y., Ma Y., Chen S., Ho T, & Chai Y. (2020). Runx2 Regulates Mouse Tooth Root Development Via Activation of WNT Inhibitor NOTUM. Journal of Bone and Mineral Research, 35(11), 2252-2264.

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Embryonic Mouse Head Explant Culture

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Embryonic mouse heads were dissected and collected in complete BGJb Medium: BGJb Medium (Thermo Fisher Scientific, 12591038) with 10% Fetal Bovine Serum (12662029, Thermo Fisher scientific), 0.1 mg/ml L-ascorbic acid (A4403, Sigma-Aldrich), and 1% Penicillin-Streptomycin (15140148, Thermo Fisher Scientific). Mouse heads were embedded on ice in a mixture of preheated 20% gelatin (G2500-500G, MilliporeSigma) in BGJB culture medium and sectioned to 300 μm slices using MicroSlicer Zero 1 (10111, Ted Pella). These slices were placed on cell culture inserts (PICM0RG50, Sigma Aldrich) which were inserted into 6-well culture plates and cultured at 37 °C in a 5% CO₂ incubator in complete BGJb Medium. For bead implantation, Affi-Gel blue agarose beads (1537302, BioRad) were soaked in 1 μg/μl FGF18 (100–28, Peprotech) or BSA for one hour at 37 °C (Xu et al., 2016 (link)) and then implanted into the soft palatal shelves of E14 Osr2^Cre;Tgfbr1^fl/fl head slices. The samples were collected 3 days after bead implantation.

Feng J., Han X., Yuan Y., Cho C.K., Janečková E., Guo T., Pareek S., Rahman M.S., Zheng B., Bi J., Jing J., Zhang M., Xu J., Ho T.V, & Chai Y. (2022). TGF-β signaling and Creb5 cooperatively regulate Fgf18 to control pharyngeal muscle development. eLife, 11, e80405.

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Embryonic Tissue Manipulation with Inhibitors

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Rock inhibitor (Y-27632 Calbiochem, stock: 20mM in DMSO, working: 200μM in 2% DMSO, −80°C storage), Rac1 inhibitor (NSC23766 Santa Cruz, stock: 37mM in DMSO, working: 200μM in 2% DMSO, −20°C storage), Rho inhibitor (CCG-1423 Calbiochem, stock: 22mM in DMSO, working: 200μM in 2% DMSO, −20°C storage), and JNK inhibitor (SP600125 Sigma, stock: 20mM in DMSO, working: 200μM in 2% DMSO, 4°C storage) were resuspended in DMSO, aliquoted, and stored at stock concentrations until incubation with beads. JNK activator, anisomycin (#sc-3524 Santa Cruz Biotechnology, stock 67microg/ml, working: 67ng/ml in water, 4°C storage) (Liao et al., 2006 (link)), was resuspended in water, aliquoted, and stored at 1000x stock concentrations until incubation with beads. AG 1-X2 Resin beads (Bio Rad, 140-1231, 50-100 mesh) were washed in ethanol, dried, mixed with diluted, working concentration inhibitor solution and incubated overnight at 4°C. Affi-gel blue agarose beads (50–100 mesh, Bio-Rad) loaded with anisomycin activator were prepared according to Carmona-Fontaine (2011) . Late neurula embryos had a small incision cut in their facial midline where a bead was inserted into the foregut behind the EAD. Embryos were grown to late tailbud for fixation and immunohistochemistry and to swimming tadpole for live imaging.

Jacox L., Chen J., Rothman A., Lathrop-Marshall H, & Sive H. (2016). Formation of a ‘pre-mouth array’ from the Extreme Anterior Domain is directed by neural crest and Wnt/PCP signaling. Cell reports, 16(5), 1445-1455.

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