Ubiquitin Antibody (YA022)

Ubiquitin Antibody (YA022) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to Ubiquitin.

UBB protein, a pivotal player in cellular regulation, exists either covalently attached to other proteins or in a free, unanchored state. In its covalently bound forms, UBB conjugates to target proteins through isopeptide bonds, presenting as monoubiquitin, polyubiquitin chains linked via different Lys residues, or linear polyubiquitin chains initiated at the Met residue. The functions of polyubiquitin chains are diverse and dependent on the specific Lys residue involved: Lys-6-linked ubiquitin may contribute to DNA repair, Lys-11-linked is implicated in endoplasmic reticulum-associated degradation (ERAD) and cell-cycle regulation, Lys-29-linked is associated with proteotoxic stress response and cell cycle, Lys-33-linked is engaged in kinase modification, Lys-48-linked is crucial for protein degradation via the proteasome, and Lys-63-linked plays roles in endocytosis, DNA-damage responses, and NF-kappa-B activation. Linear polyubiquitin chains, initiated at Met, are linked to cell signaling. While UBB typically conjugates to Lys residues, rare instances involve conjugation to Cys or Ser residues. In its unanchored state, free polyubiquitin contributes to distinct roles, such as activating protein kinases and participating in signaling processes. Furthermore, UBB interacts with SKP1-KMD2A and SKP1-KMD2B complexes, emphasizing its involvement in specific cellular pathways and protein interactions.

Free

P0CG47

Observed band size: 8-300 kDa

Protein A affinity purified.

Cytoplasm, Nucleus, Membrane, Mitochondrion.

Non-conjugated

Unmodified

AB_3102107

Neuroscience

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:500 ;ICC/IF: 1:50-1:200 ;IHC-P: 1:200-1:500 ;FC: 1:50-1:100

• 
  Western blot analysis of extracts from NIH3T3 (lane 2(20μg), Hela (lane 3(20μg) and HEK293T (lane 4(20μg) using Ubiquitin (HY-P80363) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of A549 cells labeling Ubiquitin with Ubiquitin Antibody (HY-P80363)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Ubiquitin Antibody (HY-P80363) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of HepG2 cells labeling Ubiquitin with Ubiquitin Antibody (HY-P80363) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Ubiquitin Antibody (HY-P80363) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse esophagus tissue using Ubiquitin Antibody (YA022). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80363, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse esophagus tissue using Ubiquitin Antibody (YA022). The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80363, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, ICC/IF, IHC-P, FC

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human Ubiquitin.AA range:90-120.