Ubiquitin-activating Enzyme/UBE1 Antibody - BSA Free

Western Blot: Ubiquitin-activating Enzyme/UBE1 Antibody [NB600-472] - Shows detection of a band at ~118 kDa corresponding to UBE1 (lane 1 800 nm channel). Approximately 35 ug of an A431 whole cell lysate was separated on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1:1,000. Incubation was for 2 h at room temperature followed by washes and reaction with a 1:10,000 dilution of IRDye (TM) 800 conjugated Gt-a-Rabbit IgG [H&L] MX10 for 45 min at room temperature. Molecular weight markers are shown in lane 2 (700 nm channel).

Immunohistochemistry: Ubiquitin-activating Enzyme/UBE1 Antibody [NB600-472] - Used at a 10 ug/ml to detect UBE1 in a variety of tissues including adrenal, breast, colon (epithelium), kidney, liver, lung (respiratory epithelium), ovary (oocyte and endothelium), pancreas (islet and exocrine), placenta, prostate (epithelium), skin (epithelium), spleen (lymphocytes), stomach (chief), testis, thymus, tonsil, and uterus (glandular, stroma). In many cells a punctate nuclear staining was observed. Other cells showed both cytoplasmic and nuclear staining. This image shows UBE1 staining of human lung tissue. Tissue was formalin-fixed and paraffin embedded. Personal Communication, Tina Roush, LifeSpanBiosciences, Seattle, WA.

Affinity Purified anti-Ubiquitin Activating Enzyme antibody was used at a 10 ug/ml to detect UBE1 in a variety of tissues including adrenal, breast, colon (epithelium), kidney, liver, lung (respiratory epithelium), ovary (oocyte and endothelium), pancreas (islet and exocrine), placenta, prostate (epithelium), skin (epithelium), spleen (lymphocytes), stomach (chief), testis, thymus, tonsil, and uterus (glandular, stroma). In many cells a punctate nuclear staining was observed. Other cells showed both cytoplasmic and nuclear staining. This image shows UBE1 staining of human lung tissue. Tissue was formalin-fixed and paraffin embedded. Personal Communication, Tina Roush, LifeSpan Biosciences, Seattle, WA.

• Reactivity: Hu, Mu, Rt, Bv, Rb
• Applications: WB, ELISA, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free

Ubiquitin-activating Enzyme/UBE1 Antibody - BSA Free Summary

• Description: The product was protein A purified from monospecific antiserum followed by further purification to remove the GST tag
  
  Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.
• Immunogen: Ubiquitin-activating Enzyme/UBE1 antibody was prepared from whole rabbit serum produced by repeated immunizations with a recombinant protein corresponding to full length Human Ubiquitin-activating Enzyme/UBE1. (Uniprot: P22314)
• Predicted Species: Mouse (96%), Rat (96%), Bovine (97%), Rabbit (97%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: UBA1
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:2000-1:10000
  • Immunohistochemistry 2 mg/ml-20 ug/ml
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Western Blot 1:1000-1:5000
• Application Notes: This purified antibody has been tested for use in ELISA, immunohistochemistry and western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band at ~118 kDa in size corresponding to UBE1 by western blotting in the appropriate cell lysate or extract.
• Control:

  Ubiquitin-activating Enzyme/UBE1 Overexpression Lysate

Reactivity Notes

A BLAST analysis was used to suggest that this antibody would react with Ubiquitin-activating Enzyme/UBE1 protein from human (100%) rabbit (96%), mouse (95%), rat (95%) and dog (93%) based on a high degree of sequence homology. Cross reactivity against this protein from other sources has not been determined.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Protein A purified

Alternate Names for Ubiquitin-activating Enzyme/UBE1 Antibody - BSA Free

• A1S9T and BN75 temperature sensitivity complementing
• A1S9T
• A1ST
• AMCX1
• GXP1
• MGC4781
• POC20 centriolar protein homolog
• POC20
• Protein A1S9
• SMAX2
• UBA1
• UBA1, ubiquitin-activating enzyme E1 homolog A
• UBA1A
• UBE1
• UBE1A1S9
• UBE1X
• ubiquitin-activating enzyme E1 (A1S9T and BN75 temperature sensitivitycomplementing)
• Ubiquitin-activating enzyme E1
• Ubiquitinactivating Enzyme
• Ubiquitin-activating Enzyme
• ubiquitin-like modifier activating enzyme 1
• ubiquitin-like modifier-activating enzyme 1

Background

Ubiquitinylation, the covalent attachment of ubiquitin to protein, regulates a number of cellular processes including responses to stress. This process is responsible for selective targeting of proteins for degradation. The first step in ubiquitinylation is the activation of ubiquitin by ubiquitin activating enzyme E1 which is a rate-limiting enzyme in the conjugation process. The ubiquitin activating enzyme exists as two isoforms: E1a (117 kDa), localized predominantly in the nucleus, and E1b (110 kDa), localized in the cytoplasm. The first 11 amino acids are considered to be essential for phosphorylation and exclusive nuclear localization of the enzyme.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Ubiquitin-activating Enzyme/UBE1 Antibody (NB600-472)(1)

Publication using NB600-472

• Panigrahi AK, Zhang N, Otta SK, Pati D. A cohesin-RAD21 interactome. Biochem J 2012-03-15 [PMID: 22145905] (Human)

Bioinformatics

• Gene Symbol: UBA1
• Entrez:
  • 7317(Human)
• Uniprot:
  • P22314(Human)