Recombinant Mouse TSLP Protein, CF

R&D Systems | Catalog # 555-TSB

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Mouse TSLP Protein (555-TSB)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

Sf 21 (baculovirus)

Accession Number

Q9JIE6

Applications

Enzyme Activity
View all TSLP Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived mouse TSLP protein
Tyr20-Glu140, with a C-terminal 10-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr20

Predicted Molecular Mass

15 kDa

SDS-PAGE

18-22 kDa triplet, reducing conditions
17 kDa after deglycosylation, reducing conditions

Activity

Used as a substrate for the endoglycosidase rFmEndo F3 (Catalog # 5548-GH).

Formulation, Preparation, and Storage

555-TSB
Formulation Supplied as a 0.2 μm filtered solution in PBS.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: TSLP

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).

References

  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.

Long Name

Thymic Stromal Lymphopoietin

Alternate Names

thymic stromal lymphopoietin

Entrez Gene IDs

85480 (Human); 53603 (Mouse); 102119254 (Cynomolgus Monkey)

Gene Symbol

TSLP

UniProt

Q9JIE6

Additional TSLP Products

Product Documents for Recombinant Mouse TSLP Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Mouse TSLP Protein, CF

For research use only

Citations for Recombinant Mouse TSLP Protein, CF

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Protocols

View specific protocols for Recombinant Mouse TSLP Protein, CF (555-TSB):

Materials
  • 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

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