Active Recombinant Human JAG1, Fc tagged

• Cat.No.: JAG1-3138H
• Product Overview: Recombinant Human JAG1 (NP_000205.1) extracellular domain (Met 1-Ser 1046), fused with the Fc region of human IgG1 at the C-terminus, was produced in Human Cell.

Specification

• Species: Human
• Source: HEK293
• Tag: Fc
• Protein Length: Met 1-Ser 1046
• Form: Lyophilized from sterile PBS, pH 7.4
• Bio-activity: Measured by the ability of the immobilized protein to enhance BMP2-induced alkaline phosphatase activity in C3H10T1/2 mouse embryonic fibroblast cells.
  The ED50 for this effect is typically 5-30 μg/mL.
• Molecular Mass: The recombinant human JAG1/Fc is a disulfide-linked homodimer. The reduced monomer consists of 1254 amino acids and has a predicted molecular mass of 137 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of human JAG1/Fc monomer is approximately 185.5 kDa due to glycosylation.
• Endotoxin: < 1.0 EU per μg protein as determined by the LAL method.
• Purity: > 85 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C
  Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.
• Publications:
  A non-canonical JAGGED1 signal to JAK2 mediates osteoblast commitment in cranial neural crest cells (2019)
  The Special Role of JAK/STAT, and Notch Signaling Pathways in Cancer Pathogenesis (2023)
  Delivery of A Jagged1-PEG-MAL hydrogel with Pediatric Human Bone Cells Regenerates Critically-Sized Craniofacial Bone Defects (2023)
  Controlled JAGGED1 delivery induces human embryonic palate mesenchymal cells to form osteoblasts (2017)
  JAGGED1 stimulates cranial neural crest cell osteoblast commitment pathways and bone regeneration independent of canonical NOTCH signaling (2021)
  Osteoinductive effect of soluble transforming growth factor beta receptor 3 on human osteoblast lineage (2021)

Gene Information

• Gene Name: JAG1 jagged 1 [ Homo sapiens ]
• Official Symbol: JAG1
• Synonyms: JAG1; jagged 1; AGS, Alagille syndrome , JAGL1; protein jagged-1; AHD; AWS; CD339; HJ1; AGS; JAGL1; MGC104644
• Gene ID: 182
• mRNA Refseq: NM_000214
• Protein Refseq: NP_000205
• UniProt ID: P78504

Citation

JAGGED1 Stimulates Cranial Neural Crest Cell Osteoblast Commitment Pathways and Bone Regeneration Independent of Canonical NOTCH Signaling

Journal: Bone    PubMed ID: 32980561    Data: 2022/4/24

Authors: Archana Kamalakar, Jay M. McKinney, Steven L. Goudy

Article Snippet:50 μL (1.5 mg) of Dynabeads Protein G ( 36 ) (Invitrogen 10003D) were transferred to a tube, where the beads were separated from the solution using a magnet.50 μL (1.5 mg) of Dynabeads Protein G ( 36 ) (Invitrogen 10003D) were transferred to a tube, where the beads were separated from the solution using a magnet.. Recombinant JAG1-Fc (5 μM, 5.7 μM, 10 μM or 20 μM) (Creative BioMart, JAG1–3138H) and IgG-Fc fragment (5 μM or 5.7 μM) (Abcam, ab90285) alone were diluted in 200 μL PBS with 0.1% Tween-20 (Fisher, BP337–500) and then added to the dynabeads.. The beads plus proteins were incubated at 4°C with rotation for 16 hours.The beads plus proteins were incubated at 4°C with rotation for 16 hours.

As a proof of concept experiment, we (A) incorporated JAG1-dynabeads complex (5 μM, 10 μM or 20 μM), dynabeads alone and BMP2 (2.5 μM) in 4% PEG-MAL hydrogels and implanted them into (B-G) 3.5 mm critical-sized defects in the parietal bones of 6–8-week old C57BL/6 mice and delivered them (J) without or (K) with CNC cells into the defects as 3 separate doses (Initial dose, Week 4, Week 8), as shown in H. After 12 weeks, we quantified differences in regenerated bone volume (J-K) within the defect and compared them between experimental groups by μCT analysis. μCT reconstructions of defects are shown in I. Data were subjected to ANOVA and Tukey’s post-test and are presented as mean (n ≥ 2) ± SD with p values reported.

(A) A principal component analysis of all treatments revealed that PC1 distinguished JAG1-treated cells to the right from non-JAG1 treated cells to the left along the horizontal axis. PC2 distinguished cells treated with JAG1 alone towards the top and JAG1 + DAPT treated cells towards the bottom along the vertical axis. (B) Clustering analysis using Pearson correlation-coefficient reveals the heatmap showing distinct gene clusters downstream of different treatments (n = 3). Cluster A and B consists of genes downregulated by JAG1 and Cluster C consists of genes upregulated by JAG1.

(A-B) Volcano plots of differentially expressed genes in CNC cells that were treated with JAG1-dynabeads complex (5.7 μM) with or without DAPT (15 μM) compared to Fc-dynabeads complex (5.7 μM). (C) Subsequent comparison of the JAG1 and JAG1 + DAPT treatment groups for genes that were significantly upregulated in either group compared to Fc. Plots indicate genes that were down-regulated in response to DAPT (red) or preserved between groups (grey). (D) Comparison between JAG1 treated-samples in the presence and absence of DAPT revealed various distinct genes, like Cxcl1, Cxcl12, Hes1 and Il6 upregulated downstream of JAG1 alone and Prl2c2 upregulated downstream of JAG1 + DAPT along with some conserved genes like Id1 and Rhou. Data were subjected to ANOVA and Tukey’s post-test and are presented as mean (n ≥ 2) ± SD with p values reported.

Controlled JAGGED1 delivery induces human embryonic palate mesenchymal cells to form osteoblasts

Journal: Journal of biomedical materials research. Part A    PubMed ID: 28913955    Data: 2019/2/1

Authors: Jean De La Croix Ndong, Yvonne Stephenson, Steven Goudy

Article Snippet:Culture well chambered coverglass were pre-coated with rabbit anti-human IgG (10 μg/mL) in phosphate saline buffer (PBS) for 30 min at 37°C and subsequently blocked with cell culture growth medium for 30 min. Chambered coverglass were then coated with 5 μg/mL JAGGED1/Fc (JAG1–3138 H, Creative BioMart) diluted in growth media for 2 hours at 37°C.. As control for JAGGED1, human IgG (5 μg/mL) was used.As control for JAGGED1, human IgG (5 μg/mL) was used.

JAGGED1 release profile and Notch signaling activation. (A) Prefunctionalized PEG-MAL with JAGGED1 (10 ug/m) and RGD peptide were cross-linked and incubated for 20 days in PBS. Released JAGGED1 from PEG-MAL collected every 2–3 days and quantified by ELISA. (B–C). HEPM cells were encapsulated in functionalized JAGGED1–5%PEG-MAL hydrogels and cultured in growth media for 3 days. HEPM cells proliferation (B) and Apoptosis were measured using the CellTiter Aqueous One 96 Kit and Apo-One Homogeneous Caspase3/7 kit, respectively. (n = 3) *p <0.05 versus IgG control (JAGGED1 (0 ug/mL) and #p <0.05 versus day 0.

Activation of Notch Signaling Pathway by Jagged-1PEG-MAL. HEPM cells or Notch reporter CHO cells were encapsulated in functionalized JAGGED1–5%PEG-MAL hydrogels and cultured for 3 days. (A) Notch signaling activation in CHO cells analyzed by YFP fluorescence and quantified using Image J. (bar: 100 μm). (B) Notch-associated gene expression in HEPM cells were assessed by RT-PCR. (n = 3) *p <0.05 versus IgG control (JAGGED1 (0 μg/mL).

JAGGED1-PEG-MAL stimulates the differentiation of HEPM cells in vitro. HEPM cells were encapsulated in functionalized JAGGED1–5%PEG-MAL hydrogels and cultured in osteogenic media for 7 days. (A) HEPM differentiation analyzed by ALP activity using p-nitrophenyl phosphate (pNPP) as a substrate which turns yellow when dephosphorylated by ALP. (B) Expression of osteoblast marker genes was assessed by qPCR (n = 3) *p <0.05 versus IgG control (JAGGED1 (0 μg/mL).

Reviews

07/31/2024

JAG1-3138H

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