Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [2]
- Other assay [2]
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- Product number
- PA5-14542 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Cullin 4A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with mouse based on sequence homology.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 400 µL
- Concentration
- 2 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Selective inhibition of cullin 3 neddylation through covalent targeting DCN1 protects mice from acetaminophen-induced liver toxicity.
A potent small-molecule inhibitor of the DCN1-UBC12 interaction that selectively blocks cullin 3 neddylation.
Zhou H, Lu J, Chinnaswamy K, Stuckey JA, Liu L, McEachern D, Yang CY, Bernard D, Shen H, Rui L, Sun Y, Wang S
Nature communications 2021 May 11;12(1):2621
Nature communications 2021 May 11;12(1):2621
A potent small-molecule inhibitor of the DCN1-UBC12 interaction that selectively blocks cullin 3 neddylation.
Zhou H, Lu J, Liu L, Bernard D, Yang CY, Fernandez-Salas E, Chinnaswamy K, Layton S, Stuckey J, Yu Q, Zhou W, Pan Z, Sun Y, Wang S
Nature communications 2017 Oct 27;8(1):1150
Nature communications 2017 Oct 27;8(1):1150
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CUL4a (arrow) using a Cullin 4A polyclonal antibody (Product # PA5-14542) in 293 cell lysates (2 µg/lane) either nontransfected (Lane 1) or transiently transfected with the CUL4a gene (Lane 2).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of CUL4a (arrow) using a Cullin 4A polyclonal antibody (Product # PA5-14542) in 293 cell lysates (2 µg/lane) either nontransfected (Lane 1) or transiently transfected with the CUL4a gene (Lane 2).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 5 DI-591 selectively and rapidly inhibits neddylation of cullin 3. a Western blotting of neddylated and un-neddylated cullin family members and several representative well-known substrates of cullin CRLs in KYSE70 esophageal cancer cells and THLE2 immortalized liver cells. Cells were treated with DCN1 inhibitor DI-591 or NAE E1 inhibitor MLN4924 at indicated concentrations, or DI-591DD at 10 uM for 24 h. Protein levels of neddylated and un-neddylated cullin family members and several known substrates of cullin CRLs were examined by western blotting analysis. GAPDH was used as a loading control. b Inhibition kinetics of neddylation of cullin 1 and 3 by DI-591 and MLN4924 in THLE2 liver cells. Cells were treated with DI-591 at 10 uM or MLN4924 at 0.3 uM at indicated time points. Protein levels of neddylated and un-neddylated cullin 1 and 3 were examined by western blotting analysis. GAPDH was used as a loading control. c qRT-PCR analysis of mRNA levels of NRF2 and NRF2-reguated genes in THLE2 cells. Cells were treated with DI-591 at 10 uM, DI-591DD at 10 uM or DMSO for indicated time points. The relative mRNA levels of NRF2, NQO1 and HO-1 were examined by quantitative real-time RT-PCR assay. GAPDH was used as an internal control. The averages and standard deviations for each column were calculated from total nine samples obtained from three independent experiments (each experiment with triplicates). P -value from t-test: * P < 0.05, ** P < 0.01. d Effect of siRNAs-mediated
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Fig. 6 DI-1548 and DI-1859 selectively inhibit cullin 3 neddylation in different types of cells. The effects of a dose range of the indicated compound on the protein level in osteosarcoma U2OS ( a ) and in immortalized THLE2 liver cell lines ( b ). The protein levels of neddylated cullin 3, 1, 2, 4A, 4B, and 5 (N-Cul3, 1, 2, 4A, 4B, and 5) and unneddylated cullin 3, 1, 2, 4A, 4B, and 5 (Cul3, 1, 2, 4A, 4B, and 5), as well as the substrates of cullins mediated CRLs, including p21, Bim, Nrf2 and CDT1, and HO-1 were examined by western blotting analysis. GAPDH was used as a loading control. c DI-1548 rapidly and selectively inhibits the neddylation of cullin 3 in osteosarcoma U2OS. The protein levels of neddylated cullin 3, 1 (N-Cul3, 1) and unneddylated cullin 3, 1 (Cul3, 1) were examined by western blotting analysis. GAPDH was used as a loading control. d DI-1548 effectively engages cellular DCN1 protein. U2OS cell line was treated by dose ranges of compounds DI-1548, DI-1548DD, or DI-591 for 1 h, then was heated at 53 degC. The cells were snap frozen in liquid nitrogen and thawed for three cycles. The levels of DCN1 and DCN3 in whole cell lysates were examined by western blotting analysis. GAPDH was used as a loading control. Representative images of two independent experiments are shown for a - d . Source data are provided as a Source Data file.