Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [3]
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Validation data
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- Product number
- 703675 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HERC5 Recombinant Rabbit Monoclonal Antibody (8H23L10)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Cat, Sheep.
- Antibody clone number
- 8H23L10
- Concentration
- 0.5 mg/mL
Submitted references YAP ISGylation increases its stability and promotes its positive regulation on PPP by stimulating 6PGL transcription.
Xue X, Tian X, Zhang C, Miao Y, Wang Y, Peng Y, Qiu S, Wang H, Cui J, Cao L, Sun F, Qiao Y, Zhang X
Cell death discovery 2022 Feb 11;8(1):59
Cell death discovery 2022 Feb 11;8(1):59
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of HERC5 was achieved by transfecting HeLa cells with HERC5 specific siRNA (Silencer® select Product # s27646 & s27647). Western blot analysis (Fig a) was performed using whole cell extracts from HERC5 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-HERC5 Recombinant Rabbit Monoclonal Antibody (Product # 703675, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this Western blot is shown in the histogram (Fig b). Loss of signal upon siRNA mediated knockdown confirms that antibody is specific to HERC5.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with IFN-alpha (500 U/ml for 48 hrs) (Lane 2), Jurkat (Lane 3) and A549 (Lane 4). The blot was probed with Anti-HERC5 Recombinant Rabbit Monoclonal Antibody (Product # 703675, 1:500 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution). A ~117 kDa band corresponding to HERC5 was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoblot with monoclonal HERC5 antibody (Product # 703675). NK-92, HeLa, Jurkat, and A549 cells were either untreated or IFN-b-treated for 48 hours as indicated. 8220Transfected8221 (last lane) represents extract from HEK293T cells transfected with plasmids expressing 3X-FLAG-ISG15, UBE1L, UBCH8, and HERC5. Cells were lysed in 1% NP40 lysis buffer (50 mM Tris pH 8, 150 mM NaCl, 1% NP-40) with PMSF, aprotinin, leupeptin, and 1mM DTT. 30 mg total protein per sample were loaded onto a 13% Tris-Glycine PAGE gel, transferred to nitrocellulose using a semi-dry transfer apparatus (0.1A for 1 hour 15 minutes), and blocked for 30 minutes at RT. The membrane was probed with anti-ISG15 (1:1000 in 1X TNET) for one hour at RT, washed with 1X TNET, and incubated with secondary antibody (1:15,000) for one hour at RT. Protein marker used is Thermo Page Ruler Plus (Product # 26620). Data courtesy of Caleb Swaim and Jon Huibregtse at University of Texas at Austin.