Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Other assay [3]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA1-187 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Ubiquitin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- PA1-187 detects both the ubiquitin monomer as well as poly-ubiquitinated proteins of various molecular weights.
- Reactivity
- Human, Mouse, Rat, Canine
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- -20°C
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Ubiquitin was performed by loading 20 µg of the indicated whole cell lysates per well, and 10 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 1 hour at room temperature. Ubiquitin was detected at ~8.5 kDa after probing with a Ubiquitin polyclonal antibody (Product # PA1-187) at a dilution of 1:1000 in StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) overnight at 4C on a rocking platform, washing in TBST, and probing with an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: In addition to detecting ubiquitin monomer, this antibody also detects poly-ubiquitinated proteins at various molecular weights.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Ubiquitin was performed on HeLa cells either left untreated (right panel) or treated with 10uM of MG132 proteasome inhibitor (left panel) for 2 hours at 37C. Whole cell lysates were prepared using IP Lysis buffer (Product # 87787), and 10 µg of the lysates along with 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) were loaded onto a Novex® 10-20% Tricine Gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288) and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for 1 hour at room temperature. Ubiquitin was detected at ~8.5 kDa after probing with a Ubiquitin polyclonal antibody (Product # PA1-187) at a dilution of 1:1000 in StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) overnight at 4C on a rocking platform, washing in TBST, and probing with an HRP-conjugated goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: In addition to detecting ubiquitin monomer, this antibody also detects poly-ubiquitinated proteins at various molecular weights.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3. Nuclear protein quality control degrades non-imported mitochondrial proteins. ( A ) Western blot of yeast expressing Ilv2-GFP +- FCCP +- MG-132. ( B ) Western blot of yeast expressing Ilv2-GFP +- FCCP in wild-type (WT) and E3 KO strains. ( C ) Western blots showing cycloheximide (CHX) chase of Ilv2-GFP in WT and E3 KO strains in the presence of FCCP. ( D ) Western blot showing ubiquitylation of immunoprecipitated Ilv2-GFP +- FCCP in WT and E3 KO strains. Pgk1 = loading control. E3 KO = san1 Delta ubr1 Delta doa10 Delta. P = precursor and M = mature. Figure 3--figure supplement 1. Nuclear protein quality control promotes non-imported mitochondrial protein degradation. ( A ) Western blot of yeast expressing Ilv2-HA +- FCCP +- MG-132. ( B, C ) Western blots of yeast expressing the Ilv2-HA ( B ) or endogenous Ilv2 ( C ) +- FCCP in WT and E3 KO strains. ( D ) Western blots of yeast expressing Ilv2-GFP +- FCCP in WT and the indicated mutant yeast strains. ( E, F ) Western blots showing the CHX chase of Ilv2-HA ( E ) or endogenous Ilv2 ( F ) in WT and E3 KO strains in the presence of FCCP. ( G ) Western blots of yeast expressing the Lat1-GFP +- FCCP in WT and E3 KO strains. P = precursor form, M = mature form. Pgk1 = loading control. Figure 3--figure supplement 2. Nuclear protein quality control promotes non-imported mitochondrial protein degradation. ( A ) Yeast expressing the indicated GFP and mCherry tagged mitochondrial proteins +- FCCP. ( B ) Quantification of ( A ).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 p97-R155C is functionally defective for chromatin extraction. ( A ) pDNA was incubated in extract supplemented with p97-WT, p97-D1D2, or p97-R155C. DNA-bound proteins were isolated by plasmid pull-down at various times and visualized by Western blot with the indicated antibodies. Note that p97 antibodies recognize both endogenous and recombinant proteins. ( B , C ) Relative quantitation of total DNA-bound p97 ( B ) and ubiquitin ( C ) from three biological replicates. Values are normalized to peak accumulation in the +p97-WT reaction and error bars represent +/- one standard deviation. Full Western blot images are available in Fig. S7 .