Recombinant Human Hyaluronidase 2/HYAL2 His-tag Protein, CF

R&D Systems | Catalog # 11012-GH

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Key Product Details

  • R&D Systems CHO-derived Recombinant Human Hyaluronidase 2/HYAL2 His-tag Protein (11012-GH)
  • 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

CHO

Accession Number

Q12891.4

Applications

Enzyme Activity
View all Hyaluronidase 2/HYAL2 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human Hyaluronidase 2/HYAL2 protein
Met21-Gly447, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Met21

Predicted Molecular Mass

50 kDa

SDS-PAGE

53-58 kDa, under reducing conditions.

Activity

Measured by its ability to digest Cy5-Labeled Hyaluronan (Low MW) to dp1-dp4.
The majority of Cy5-Labeled Hyaluronan (Low MW) (2 pmol) is digested by 0.25 μg of Recombinant Human HYAL-2, as measured under the described conditions.

Scientific Data Images for Recombinant Human Hyaluronidase 2/HYAL2 His-tag Protein, CF

Recombinant Human HYAL-2 His-tag Protein  (rhHYAL-2) Enzyme Activity Diagram

rhHYAL-2 recognizes repeating subunits of -GlcA-beta 1,­3-GlcNAc-beta 1,4- and hydrolyzes high molecular weight hyaluronic acid to intermediate and smaller sized products. 

Recombinant Human Hyaluronidase 2/HYAL2 SDS-PAGE

1 μg/lane of rhHYAL-2 (Catalog # 11012-GH) was resolved with SDS-PAGE under reducing (R) conditions and visualized by Silver Staining, showing a band at 55 kDa.

Fluorescent Gel Mobility Shift caused by rhHYAL-2.

Lane 1 contained substrate Cy5-labeled LMW-HA GL401. In the presence of rhHYAL-2, the Hyaluronic Acid was modified to smaller molecular weight products with increasing amounts of rhHYAL-2 from left to right and a mobility shift was observed.

Formulation, Preparation, and Storage

11012-GH
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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: Hyaluronidase 2/HYAL2

Human hyaluronidases (HYALs) are a group of five endo­-beta -­N­-acetyl­hexosaminidases that include HYAL1, HYAL2, HYAL3, HYAL4, and SPAM1 (PH20) (1, 2). While HYAL1, HYAL2 and SPAM1 are mainly active on hyaluronan (3, 4, 5), HYAL4 is active on both hyaluronan (6) and chondroitin sulfate (type C and D in particular) (7). Hyaluronan and chondroitin sulfate are abundant extracellular matrix components that have numerous biological functions (8). The backbone of chondroitin sulfate composed of repeating units of ­4-GlcA-beta 1,­3-GalNAc-beta 1­ most closely resembles to that of hyaluronan composed of repeating units of ­4-GlcA-beta 1,­3-GlcNAc-beta 1­. While HYAL1 is a strict lysosomal hyaluronidase and is active when pH ≤ 5.5 (9), HYAL2 is active in a narrow range of pH 5.0 to 5.5 (6) and is attached to cell membrane via GPI linker (10) where they may serve to hydrolyze high molecular weight hyaluronic acid of extracellular matrix to intermediate and small sized products.

References

  1. Csoka, A.B. et al. (2001) Matrix Biol. 20:499.
  2. Jedrzejas, M. J. and Stern, R. (2005) Proteins 61:227.
  3. Csoka, A.B. et al. (1999) Genomics 60:351.
  4. Lin, Y. et al. (1994) J. Cell Biol.125:1157.
  5. Lepperdinger, G. et al. (2001) Matrix Biol. 20:509.
  6. Wu, Z.L. and Ertelt, J.M. (2021) Glycobiology, cwab061, https://doi.org/10.1093/glycob/cwab061.
  7. Kaneiwa, T. et al. (2010) Glycobiology 20:300.
  8. Iozzo, R.V. (1998) Annu. Rev. Biochem. 67:609.
  9. Hofinger, E.S.A. et al. (2007) Glycobiology 17:444.
  10. Rai, S.K. et al. (2001). Proc. Natl. Acad. Sci. U.S.A, 98:4443.

Long Name

Hyaluronoglucosaminidase-2

Alternate Names

HYAL-2, LUCA-2, LUCA2

Entrez Gene IDs

8692 (Human)

Gene Symbol

HYAL2

UniProt

Q12891.4

Additional Hyaluronidase 2/HYAL2 Products

Product Documents for Recombinant Human Hyaluronidase 2/HYAL2 His-tag Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human Hyaluronidase 2/HYAL2 His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human Hyaluronidase 2/HYAL2 His-tag Protein, CF (11012-GH):

Materials
  • Assay Buffer: 50 mM MES, pH 5.0
  • Recombinant Human HYAL-2 (rhHYAL-2) (Catalog # 11012-GH)
  • Cy5-Labeled Hyaluronan (Low MW) (Cy5 LMW-HA) (Catalog # GL401)
  • 17% SDS-PAGE gel and SDS-PAGE Reagents
  • Reducing SDS-PAGE sample buffer
  • Fluorescent imager
  1. Dilute rhHYAL-2 to 25 ng/µL in Assay Buffer.
  2. Dilute Cy5 LMW-HA to 0.2 µM in Assay Buffer.
  3. Prepare reaction by combining 10 µL of diluted rhHYAL-2 and 10 µL of diluted Cy5 LMW-HA.
  4. Prepare a negative control by combining 10 µL of diluted Cy5 LMW-HA and 10 µL of Assay Buffer.
  5. Incubate reaction(s) and negative control at room temperature for 18 hours.
  6. Add 7 µL of the reducing SDS-PAGE sample buffer to each reaction and negative control.
  7. Load 13.5 µL each sample and control per well on a 17% SDS-PAGE gel and run SDS-PAGE until the dye front has migrated more than two thirds of the way down the gel.
  8. Acquire gel image with a fluorescent imager.
  9. Visually determine the digestion of Cy5 LMW-HA by rhHYAL-2 to dp 1-4 in each reaction lane.

Per Reaction:

  • rhHYAL-2: 0.25 µg
  • Cy5 LMW-HA: 2 pmol

FAQs

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