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Recombinant Mouse TSLP Protein, CF

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Product Details

Summary
Reactivity MuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

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Recombinant Mouse TSLP Protein, CF Summary

Details of Functionality
Used as a substrate for the endoglycosidase rFmEndo F3 (Catalog # 5548-GH).
Source
Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse TSLP protein
Tyr20-Glu140, with a C-terminal 10-His tag
Accession #
N-terminal Sequence
Tyr20
Protein/Peptide Type
Recombinant Enzymes
Gene
Tslp
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Dilutions
  • Enzyme Activity
Theoretical MW
15 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
18-22 kDa triplet, reducing conditions
17 kDa after deglycosylation, reducing conditions
Publications
Read Publication using
555-TSB in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in PBS.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Assay Procedure
  • Assay Buffer: 50 mM NaOAc, pH 4.5
  • Recombinant F. meningosepticum Endo-beta -N-acetylglucosaminidase F3/Endo F3 (rFmEndo F3) (Catalog # 5548-GH)
  • Substrate: Recombinant Mouse TSLP (rmTSLP) (Catalog # 555-TSB)
  • 15% SDS-PAGE gel
  • Reducing SDS-PAGE gel loading buffer (≥3X concentration)
  • Silver Staining reagents
  • BioRad GS-800 densitometer (or equivalent)
  1. Dilute rFmEndo F3 to 5, 2.5, 1.25, 0.625. 0.313, 0.156, 0.078 μg/mL in Assay Buffer.
  2. Dilute Substrate to 100 μg/mL in Assay Buffer.
  3. Combine 10 μL rFmEndo F3 at each dilution with 10 μL of 100 μg/mL Substrate. Include a control containing 10 μL Assay Buffer and 10 μL of 100 μg/mL Substrate.
  4. Incubate the reaction and control at 37 °C for 30 minutes.
  5. Add 10 μL of reducing gel buffer to each reaction. Boil sample at 100 °C for 3 to 5 minutes before loading to gel.
  6. Load all of the sample (30 μL) per lane on a 15% gel. As a staining development control load 25 ng BSA.
  7. Perform electrophoresis.
  8. Stain gel with silver stain, stop stain development when 25 ng BSA becomes visible.
  9. Analyze % deglycosylation by densitometry.
  10. Determine the DC50 for rFmEndoF3 by plotting % Substrate deglycosylated vs. amount with 4-PL fitting.
Per Lane:
  • rFmEndo F3: 50, 25, 12.5, 6.25, 3.13, 1.56, 0.78 ng
  • rmTSLP: 1 μg

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Mouse TSLP Protein, CF

  • thymic stromal lymphopoietin
  • TSLP

Background

Stromal Lymphopoietin (TSLP) was originally identified from the conditioned medium of a mouse thymic stromal cell line as a protein that promoted the development of B cells. The activity of mouse TSLP overlaps with, but is distinct from, that of mouse IL-7 (1). Mouse TSLP cDNA encodes a 140 amino acid (aa) residue precursor protein with a 19 aa signal sequence. Within the mature region, there are three potential N-glycosylation sites. The Sf 21 cell expressed recombinant mouseTSLP is likely to be glycosylated at all three sites, as three major glycoforms were visible on SDS-PAGE (Figure 1). Insect cells are known to express relatively simple and homogeneous N-glycans that mainly belong to the high mannose type (2). This recombinant protein was found to be an excellent substrate for N-specific glycosidases such as Endo F3 (Figure 1). The majority of the glycans on recombinant mouse TSLP can be readily removed by Endo F3. However, a small percentage of the glycans is somewhat resistant to Endo F3 digestion, possibly lacking core fucose, as it is known that core fucosylated N-glycans are strongly preferred substrates for Endo F3 digestion (3).

  1. Sims, J.E. et al. (2000) J. Exp. Med. 192:671.
  2. Staudacher, E. et al. (1992) Eur. J. Biochem. 207:987.
  3. Tarentino, A.L. and T.H. Jr. Plummer (1994) Glycobiology 4: 771.

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Publications for TSLP (555-TSB)(1)

We have publications tested in 1 confirmed species: Mouse.

We have publications tested in 1 application: In Vivo.


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Bioinformatics

Gene Symbol Tslp
Uniprot