Human UBE2I/Ubc9 Antibody (1050001)

R&D Systems | Catalog # MAB11185

R&D Systems

Key Product Details

Species Reactivity

Human

Applications

Western Blot, Immunocytochemistry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 1050001
View all UBE2I/Ubc9 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived human UBE2I/Ubc9 protein
Met1-Ser158
Accession # P63279

Specificity

Detects Human UBE2I/Ubc9 in direct Elisa.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Scientific Data Images for Human UBE2I/Ubc9 Antibody (1050001)

Detection of Human UBE2I/Ubc9 by Western Blot.

Western blot shows lysates of U937 human histiocytic lymphoma cell line, HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human UBE2I/Ubc9 Monoclonal Antibody (Catalog # MAB11185) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). A specific band was detected for UBE2I/Ubc9 at approximately 18 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Detection of UBE2I/Ubc9 in A431 Human Epithelial Carcinoma Cell Line.

UBE2I/Ubc9 was detected in immersion fixed A431 cells using Mouse Anti-Human UBE2I/Ubc9 Monoclonal Antibody (Catalog # MAB11185) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to Nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Human UBE2I/Ubc9 Antibody (1050001)

Application
Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed A431 cells

Western Blot

2 µg/mL
Sample: U937 human histiocytic lymphoma cell line, HUVEC human umbilical vein endothelial cells

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: UBE2I/Ubc9

Ubiquitin-conjugating Enzyme E2I (UBE2I), also known as Ubiquitin-conjugating Enzyme 9 (Ubc9), is a ubiquitously expressed protein with a predicted molecular weight of 20 kDa. Human UBE2I/Ubc9 shares 100% amino acid sequence identity with the mouse and rat orthologs. UBE2I/Ubc9 catalyzes the addition of the Ubiquitin-like protein SUMO to target proteins (1,2). SUMO is transferred from a Ubiquitin-like activating (E1) enzyme heterodimer consisting of SAE1 and UBA2 to Cys93 of UBE2I/Ubc9 (3). In contrast to most Ubiquitin-conjugating (E2) enzymes that function in a complex with Ubiquitin ligases (E3s), UBE2I/Ubc9 can interact directly with protein substrates and may also play a role in substrate recognition (4). UBE2I/Ubc9 mediates the SUMOylation of a variety of proteins including RanGAP1, HDAC4, and PML (5,6). Post-translational modifications of UBE2I/Ubc9, such as auto-SUMOylation at Lys14 or Ser71 phosphorylation by CDC2/CDK1, alter UBE2I/Ubc9 catalytic activity and target protein recognition (5,7). UBE2I/Ubc9-dependent SUMOylation reduces the levels of the stem cell marker Nanog, implicating it in embryonic stem cell pluripotency maintenance (8). SUMOylation also regulates the protein levels of tumor suppressors and oncogenes, and UBE2I/Ubc9 dysregulation is thought to contribute to the pathogenesis of human cancers (9).

References

  1. Desterro, J.M. et al. (1999) J. Biol. Chem. 274:10618.
  2. Johnson, E.S. & G. Blobel (1997) J. Biol. Chem. 272:26799.
  3. Johnson, E.S. et al. (1997) EMBO J. 16:5509.
  4. Yunus, A.A. & C.D. Lima (2006) Nat. Struct. Mol. Biol. 13:491.
  5. Knipscheer, P. et al. (2008) Mol. Cell 31:371.
  6. Duprez, E. et al. (1999) J. Cell Sci. 112:381.
  7. Su, Y.F. et al. (2012) PLoS One 7:e34250.
  8. Wu, Y. et al. (2012) PLoS One 7:e39606.
  9. Mo, Y.Y. et al. (2005) Oncogene 24:2677.

Long Name

Ubiquitin-conjugating Enzyme E2I

Alternate Names

p18, SUMO-protein Ligase, Ubc9, UBCE9

Entrez Gene IDs

7329 (Human); 22196 (Mouse)

Gene Symbol

UBE2I

UniProt

P63279 (Human)

Additional UBE2I/Ubc9 Products

Product Documents for Human UBE2I/Ubc9 Antibody (1050001)

Certificate of Analysis

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Manufacturing Specifications


Product Specific Notices for Human UBE2I/Ubc9 Antibody (1050001)

For research use only

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Protocols

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