Anti-FLAG M2 (F3165), anti-Myc (M4439) and anti-HA (H9658) antibodies were purchased from Sigma-Aldrich. Anti-KLHL12 (GTX83228), anti-CUL3 (GTX62065), anti-cyclin B1 (GTX100911), anti-β-catenin (GTX101435) and anti-streptavidin HRP (GTX85912) antibodies were obtained from GeneTex. Anti-actin (MAB1501) antibody was purchased from Merck Millipore. Anti-KHSRP (A302-021A) antibody was purchased from Bethyl Laboratories. Anti-PTBP1 (sc-16547) antibody was obtained from Santa Cruz Biotechnology. Anti-His (OB05) antibody was purchased from CalBioChem. Anti-PCBP2 antibody was a gift from Dr Ian Goodfellow. Anti-EV71 2B antibody was provided by Dr Jim-Tong Horng.
Anti ha h9658
Manufactured by Merck Group
Sourced in United States
Anti-HA (H9658) is a monoclonal antibody that specifically binds to the hemagglutinin (HA) epitope. It can be used for the detection and purification of HA-tagged proteins.
Automatically generated - may contain errors
Lab products found in correlation
Anti flag f1804, by Merck Group (2 mentions)
Anti actin mab1501 antibody, by Merck Group (1 mentions)
Anti ub p4d1, by Santa Cruz Biotechnology (1 mentions)
Anti mdc1, by Merck Group (1 mentions)
Anti γh2ax 05 636, by Merck Group (1 mentions)
Anti β actin a1978, by Merck Group (1 mentions)
Anti 53bp1 a300 272a, by Fortis Life Sciences (1 mentions)
Ha flag usp13, by Addgene (1 mentions)
Pgex 4t 2 vector, by Takara Bio (1 mentions)
Anti brca1 d9, by Santa Cruz Biotechnology (1 mentions)
Anti myc 9e10, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Anti e2, by Santa Cruz Biotechnology (1 mentions)
Anti myc m4439, by Merck Group (1 mentions)
Anti sir2 yn 19, by Santa Cruz Biotechnology (1 mentions)
Mouse anti flag f3165, by Merck Group (1 mentions)
Anti nemo, by Cell Signaling Technology (1 mentions)
Anti pgk1 22c5d8, by Thermo Fisher Scientific (1 mentions)
7 protocols using anti ha h9658
1
Antibody Sourcing for Protein Analysis
2
Characterization of USP13 Regulation
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HA-FLAG-USP13 was purchased from Addgene (Plasmid #22568, provided by Dr Wade Harper) and subcloned into pGEX-4 T-2 vector (Clontech). USP13 site mutants were generated by site-directed mutagenesis (Stratagene).
The anti-USP13 (GTX118595, dilution: 1:500) and anti-Rad51 (N1C2, dilution: 1:200) antibodies were purchased from Genetex. Anti-Ub (P4D1, dilution: 1:500), anti-RPA32 (9H8, dilution: 1:200) and anti-BRCA1 (D9, dilution: 1:200) antibodies were purchased from Santa Cruz Biotechnology. Anti-γH2AX (05-636, dilution: 1:500), anti-FK2 (04-263, dilution: 1:500) and anti-MDC1 (05-1572, dilution: 1:200) were purchased from Millipore. Anti-RAP80 (A303-763A, dilution: 1:500) and anti-53BP1 (A300-272A, dilution: 1:500) were purchased from Bethyl Laboratories. Anti-FLAG (F1804, dilution: 1:1,000), anti-HA (H9658, dilution: 1:1,000), and anti-β-actin (A1978, dilution: 1:2,000) antibodies were purchased from Sigma. Anti-RNF8 (ab4183, dilution: 1:500) was purchased from Abcam.
The anti-USP13 (GTX118595, dilution: 1:500) and anti-Rad51 (N1C2, dilution: 1:200) antibodies were purchased from Genetex. Anti-Ub (P4D1, dilution: 1:500), anti-RPA32 (9H8, dilution: 1:200) and anti-BRCA1 (D9, dilution: 1:200) antibodies were purchased from Santa Cruz Biotechnology. Anti-γH2AX (05-636, dilution: 1:500), anti-FK2 (04-263, dilution: 1:500) and anti-MDC1 (05-1572, dilution: 1:200) were purchased from Millipore. Anti-RAP80 (A303-763A, dilution: 1:500) and anti-53BP1 (A300-272A, dilution: 1:500) were purchased from Bethyl Laboratories. Anti-FLAG (F1804, dilution: 1:1,000), anti-HA (H9658, dilution: 1:1,000), and anti-β-actin (A1978, dilution: 1:2,000) antibodies were purchased from Sigma. Anti-RNF8 (ab4183, dilution: 1:500) was purchased from Abcam.
3
Western Blot Analysis of Protein Expression
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Cells were lysed in cell lysis solution (Pierce) on ice for 1 h and centrifuged at 12,000 rpm for 5 min to remove cell debris. The supernatants were subjected to SDS-PAGE. Proteins were then transferred to a 0.45-μm polyvinylidene difluoride (PVDF) membrane and labeled with specific antibodies. The antibodies used in this subject were as follows: anti-MYC (9E10, 1:1,000), anti-HA (H9658, 1:1,000), anti-FLAG (F1804, 1:1,000), and anti-beta-actin (A2066, 1:1,000) were from Sigma; anti-SAMHD1 (ab67820, 1:1,000), anti-NS3 (ab65407, 1:1,000), and anti-Core (ab2740, 1:1,000) were from Abcam; anti-SREBP-1 (39940, 1:1,000) was from Active Motif; HRP-conjugated goat anti-mouse (ZSGB-Bio, catalog ZB2305, 1:5,000) and goat anti-rabbit (ZSGB-Bio, catalog ZB2301, 1:5,000) were secondary antibodies.
4
HPV18 E2 Interacts with Caspase-8
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HEK293 cells were maintained in DMEM media supplemented with 10% foetal bovine serum (FBS). Cells were seeded in 6-well plates and co-transfected with 1.5 μg of pEGFPc1 HPV18 E2 and 500 ng of pcDNA3.0 caspase-8 vectors using Lipofectamine 2000 (Life Technologies), as per the manufacturer’s instructions. Post 24 h, the transfected cells were lysed and immunoprecipitated using 2 μg of anti-E2 or anti-FADD (sc-56093; Santa Cruz Biotechnology, Inc.) as described earlier22 (link). Eluted proteins and cell lysates were separated on SDS-PAGE and immunoblotted using anti-HA (H9658; Sigma-Aldrich, USA) or anti-E2 (sc-26939, Santa Cruz Biotechnology, Inc.) antibodies.
5
Immunoblotting of Yeast Proteins
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The preparation of yeast protein extract from TCA-treated cells was described previously [57 (link)]. The samples were separated by electrophoresis on a 10% SDS-polyacrylamide gel. The following primary antibodies were used for immunoblotting at a 1:1000 dilution: anti-Pgk1 (ab113678), anti-Rad53 (ab104232) and anti Y-H2A (ab15083) were obtained from Abcam; anti-Rpd3 (yC-19), anti-Hda1 (yC-20) and anti-Sir2 (yN-19) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA); and anti-HA (H9658) and anti-Myc (M4439) were obtained from Sigma-Aldrich. Anti-mouse (A9044), anti-rabbit (A0545) and anti-goat (A5420) HRP-linked secondary antibodies were obtained from Sigma-Aldrich (St. Louis, MO, USA) and used at a 1:100,000 dilution. Images were acquired with a Wealtec KETA-CL imaging system.
6
Antibody Validation for Protein Analysis
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Mouse anti-Myc (M47-3), anti-α-tubulin M175-3), anti-His (D291-3) and rabbit anti-β-actin were purchased from MBL (Nagoya, Japan); mouse anti-Flag (F3165) and anti-HA (H9658) from Sigma (St. Louis, MO, USA); and rabbit anti-IκBα (No.9242), anti-p-IκBα (No.2859), anti-p65 (No.4764), anti-USP7 (No.4833) and anti-NEMO (No.2685) from Cell Signaling Technology (Beverly, MA, USA). Recombinant TNFα and IL-1 were from PeproTech (Rocky Hill, NJ, USA). HSCARG was cloned into expression vectors pRK-HA, pRK-Flag and pcDNA-Myc-His, respectively, and verified by DNA sequencing. NEMO were cloned into pRK-HA or pRK-Flag expression vector. The shRNA plasmids were constructed by Shanghai Genechem Corporation (Shanghai, China). siRNA of NEMO (sc-29363) and IKKβ (sc-35644) were from Santa Cruz (Dallas, TX, USA). The sequences of the sense strand of USP7 shRNA was 5′-ACCCUUGGACAAUAUUCCU-3′ and 5′-AGUCGUUCAGUCGUCGUAU-3′.
7
Yap8 Protein Stability Assay
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To determine Yap8 protein stability, yeast cells were either untreated or exposed to 0.5 mM As(III). Then, protein translation was stopped by the addition of cycloheximide (CHX) to a final concentration of 50 µg/ml. Cells were harvested at the indicated time points and protein extracts were obtained using the alkali method. Similar amounts of protein were resolved on SDS–polyacrylamide gels and transferred to nitrocellulose membranes. The primary antibodies used include anti-HA (H9658, Sigma-Aldrich, dilution 1:5000) and anti-Pgk1 (22C5D8, Thermo Fisher Scientific, Waltham, MA, USA, dilution 1:10 000) and their appropriate HPR-conjugated secondary antibodies. Immunoblots were scanned using LAS-100 image reader (Fujifilm, Minato, Japan).
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