Recombinant Mouse CD40 Ligand

• Cat.No.: Cd40lg-98M
• Product Overview: Recombinant Mouse sCD40 produced in E.Coli is a non-glycosylated, Polypeptide chain containing 149 amino acids and having a molecular mass of 16409 Dalton. The sCD40 is purified by proprietary chromatographic techniques.

Specification

• Species: Mouse
• Source: E.coli
• Tag: Non
• Description: CD40L or CD154 is a membrane glycoprotein and differentiation antigen expressed on the surface of T-cells. The CD40 ligand stimulates B-cell proliferation and secretion of all immunoglobulin isotypes in the presence of cytokines. CD40 ligand has been shown to induce cytokine production and tumoricidal activity in peripheral blood monocytes. It also costimulates proliferation of activated T-cells and this is accompanied by the production of IFN-gamma, TNF-alpha, and IL2.
• Amino Acid Sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Gln-Arg-Gly-Asp.
• Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.
• Purity: Greater than 98.0% as determined by: (a) Analysis by RP-HPLC. (b) Analysis by SDS-PAGE.
• Formulation: Lyophilized from a sterile concentrated solution (1 mg/ml) with 10mM Sodium Phosphate pH=7.5.
• Solubility: It is recommended to reconstitute the lyophilized sCD40 in sterile 18 MΩ-cm H2O not less than 100 µg/ml, which can then be further diluted to other aqueous solutions.
• Biological Activity: The ED50 as determined by its ability to induce MIP-1 alpha & TNF-alpha from mouse splenocytes was found to be 0.1 µg/ml.
• Protein Content: Protein quantitation was carried out by two independent methods: 1. UV spectroscopy at 280 nm using the absorbency value of 1.1 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). 2. Analysis by RP-HPLC, using a standard solution of sCD40 as a Reference Standard.
• Storage: Lyophilized sCD40 although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution CD154 should be stored at 4°Cbetween 2-7 days and for future use below -18°C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.

Gene Information

• Gene Name: Cd40lg CD40 ligand [ Mus musculus ]
• Synonyms: Cd40lg; CD40 ligand; IGM; IMD3; Ly62; TRAP; gp39; CD154; Cd40l; HIGM1; Ly-62; T-BAM; Tnfsf5; OTTMUSP00000019349; tumor necrosis factor (ligand) superfamily, member 5
• Gene ID: 21947
• mRNA Refseq: NM_011616
• Protein Refseq: NP_035746
• UniProt ID: P27548
• Chromosome Location: X A5; X 18.0 cM
• Pathway: Allograft rejection; Asthma; Autoimmune thyroid disease; Cell adhesion molecules (CAMs); Cytokine-cytokine receptor interaction; Primary immunodeficiency; Systemic lupus erythematosus; T cell receptor signaling pathway
• Function: cytokine activity; tumor necrosis factor receptor binding

Citation

Epithelial-Cell-Derived Phospholipase A 2 Group 1B Is an Endogenous Anthelmintic

Journal: Cell Host & Microbe    PubMed ID: 29024642    Data: 2017/10/11

Authors: Lewis J. Entwistle, Victoria S. Pelly, Mark S. Wilson

Article Snippet:Adult L5 H. polygyrus worms were isolated from C57BL/6 following primary infection between days 14 and 28 using a modified Baermann apparatus.Adult L5 H. polygyrus worms were isolated from C57BL/6 following primary infection between days 14 and 28 using a modified Baermann apparatus.. Adult L5 H. polygyrus worms were treated with recombinant mouse PLA 2 g1B (Creative BioMart) or heat inactivated recombinant mouse PLA 2 g1B in EnzChek ? PLA 2 reaction buffer (Invitrogen) at room temperature for 24 hr.

PLA 2 g1B Has Direct Anthelmintic Properties (A) ATP concentration of H.p. L3 larvae after 24 hr treatment with PLA 2 g1B, n = 3. (B) Number of H.p. larvae imbedded in the small intestinal wall 5 days after infection following 24 hr treatment with PLA 2 g1B, n = 5. (C) Luminal H.p. worms in the small intestine 14 days after 1° infection following 24 hr treatment with PLA 2 g1B, n = 10 (data pooled from two independent experiments). (D) Pla2g1b ? /? or WT mice were orally infected with 200 L3 H.p. larvae on day 0 and were drug treated (Rx) on days 14 and 15. Mice were then reinfected with PLA 2 g1B-treated L3 H.p. larvae on day 35 and harvested 14 days after infection (2°). Another cohort of Pla2g1b ? /? or WT mice were orally infected with 200 L3 H.p. larvae on day 35 and harvested 14 days after infection (1°). (E) Luminal H.p. worms in the small intestine 14 days after 1° or 2° infection following 24 hr treatment with PLA 2 g1B. (F) Luminal H.p. worms in the small intestine 14 days after 1° infection following 24 hr treatment with PLA 2 g1B. (G) Luminal H.p. worms in the small intestine 14 days after 2° infection following 24 hr treatment with PLA 2 g1B. Data are represented as mean ± SEM, n = 4–5. All data re representative of at least two independent experiments. ? = p < 0.05, ??? = p < 0.001, ???? = p < 0.0001 determined using a two-way ANOVA with Sidak’s multiple comparison analysis, one-way ANOVA with Dunnett’s multiple comparison analysis, or an unpaired two-tailed t test. See also Figure S6 .

Pla2g1b-Treatment Related Changes in Lipid Abundance Relative abundances of phosphatidylethanolamine (PE) and other lipids extracted from PLA 2 g1B-treated (10 ng/μL) and control-treated (0 ng/μL) larvae. (A) Identified PEs. Features were regarded as “identified” by comparison of their precursor ion and MS/MS fragments with the LipidBlast library, as outlined in Figure S1 . The arrangement of fatty acid moieties on the glycerol backbone (i.e., whether in the sn- 1 or sn- 2 position) and the position of double bonds could not be inferred. PE 38:3 and PE 38:4 were detected as a mixture of different fatty acid moieties. (B) Putatively identified (annotated) PEs. The features could be “annotated” as PEs by comparison of peak retention time and inter-cluster mass shifts of 28 Da (CH 2 CH 2 ) and intra-cluster mass shifts of 2 Da (indicative of difference in double bond number [fatty acid saturation]) with other, identified PEs. MS/MS could not be performed due to low abundance. Data are shown as normalized intensities expressed in arbitrary units. Data are represented as mean ± SEM, n = 3. ? = p < 0.05. TIC: Total ion current. See also Figure S7 .

Murine germinal center B cells require functional Fms-like tyrosine kinase 3 signaling for IgG1 class-switch recombination

Journal: Proceedings of the National Academy of Sciences of the United States of America    PubMed ID: 26627255    Data: 2015/12/1

Authors: Mattias N. D. Svensson, Karin M. E. Andersson, Maria I. Bokarewa

Article Snippet:For in vitro induction of Stat6 phosphorylation, splenocytes were isolated at day 14 and stimulated with LPS (10 μg/mL) or LPS (10 μg/mL) and IL-4 (50 ng/mL).For in vitro induction of Stat6 phosphorylation, splenocytes were isolated at day 14 and stimulated with LPS (10 μg/mL) or LPS (10 μg/mL) and IL-4 (50 ng/mL).. For in vitro stimulation of Flt3 + B cells, WT B cells were stimulated for 48 h with 10 μg/mL LPS, washed, and incubated for 3 h without LPS before stimulation with recombinant mouse FL (rmFL) (200 ng/mL; Creative BioMart) for 15 min, 60 min, or 24 h.