Mouse anti ha h9658

Manufactured by Merck Group

Sourced in United States

The Mouse anti-HA (H9658) is a laboratory reagent used to detect the presence of the hemagglutinin (HA) epitope tag in various applications. It is a monoclonal antibody produced in mouse.

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Close spelling variants of this product

Anti-HA (475 citations) Mouse anti-HA (118 citations) H9658 (43 citations) Anti-HA (H9658) (7 citations) ), HA (H9658, (6 citations) Mouse monoclonal anti-HA (H9658, (2 citations)

6 protocols using mouse anti ha h9658

Characterization of Anti-MLL5 Antibody and Its Applications

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Anti-MLL5 rabbit polyclonal antibody was generated in-house against a peptide corresponding to aa 1,157–1,170 (Cheng et al., 2008 (link)) and was used throughout this study for Western blotting, immunoprecipitation, and immunofluorescence. Commercial primary antibodies used for Western blotting, immunoprecipitation, and immunofluorescence were rabbit anti-pericentrin (ab4448; Abcam), rabbit anti–γ-tubulin (T5192; Sigma-Aldrich), mouse anti–γ-tubulin (T3559; Sigma-Aldrich), mouse anti–α-tubulin (T9026; Sigma-Aldrich), human autoantibody against centromere (CREST; HCT-0100; ImmunoVision), mouse anti-PLK1 (sc-17783; Santa Cruz Biotechnology), goat anti-MLL5 (sc-18214 and sc-68636; Santa Cruz Biotechnology), mouse anti–FLAG M2 (F1804; Sigma-Aldrich), rabbit anti-HA (sc-805, Santa Cruz Biotechnology), mouse anti-HA (H9658; Sigma-Aldrich), and rabbit anti–centrin-2 (sc-27793-R; Santa Cruz Biotechnology). Anti–mouse IgG-HRP, anti–rabbit IgG-HRP (Life Technologies), or Alexa Fluor 488, 568, and 594 (A11001, A11008, A11011, A11014, and A11031; Invitrogen) were used as secondary antibodies for immunoblotting or immunofluorescence.

Zhao W., Liu J., Zhang X, & Deng L.W. (2016). MLL5 maintains spindle bipolarity by preventing aberrant cytosolic aggregation of PLK1. The Journal of Cell Biology, 212(7), 829-843.

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Sulforaphane and Zidovudine Treatment Protocol

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Sulforaphane (SFN) (S8044) and Zidovudine (AZT) (L0350) were bought from LKT Laboratories. The mouse anti-actin (A1978), mouse anti-Flag (F1804), and mouse anti-HA (H9658) monoclonal antibodies; the rabbit anti-Flag (F7425), rabbit anti-HA (H6908), and anti-Mouse IgG-FITC antibodies were purchased from Sigma. Rat anti-Myc (ab206486), mouse anti-Tubulin (TA503129) monoclonal antibodies, Rabbit anti-Lamin (EPR8985), Alexa Fluor 568-conjugated (ab175476) and Alexa Fluor 647-conjugated (ab190565) antibodies were purchased from Abcam (Cambridge, UK). The rabbit anti-GST (10000-0-AP) antibodies were purchased from Proteintech. DyLight 800-labeled goat anti-mouse (5230–0415) and DyLight 508 680-labeled goat anti-rabbit (5230–0403) secondary antibodies were purchased from KPL. Anti-eqNrf2, anti-eqKeap1 and anti-P26 antibodies were prepared in our laboratory. The nuclear extraction kit was purchased from Invent Biotechnologies (SC-003). Non-cytotoxic dosages were used in this study.

Wang Y., Ma G., Wang X.F., Na L., Guo X., Zhang J., Liu C., Du C., Qi T., Lin Y, & Wang X. (2022). Keap1 recognizes EIAV early accessory protein Rev to promote antiviral defense. PLoS Pathogens, 18(2), e1009986.

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Protein Interaction and Modification Analysis

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Flag-SUMO1, RGS-SENP1 and RGS-SENP1m were gifts from Jinke Cheng (Shanghai Jiao Tong University School of Medicine, Shanghai, China). HA-PKCα was purchased from Addgene. The following primary antibodies and dilutions were used: Mouse Anti-RGS (34610, 0.1 μg ml⁻¹ dilution) was from Qiagen. Mouse Anti-Flag (M2, F3165, 1:1,000), Mouse Anti-PKC (P5704, 1:500), Mouse Anti-HA (H9658, 1:2,000) and Anti-Flag M2 affinity gel were from Sigma. Rabbit Anti-SUMO1 (4940, 1:1,000), Rabbit Anti-MARCKS (1:1,000), Rabbit Anti-p-MARCKS (1:1,000) and Rabbit Anti-PKCα (1:1,000) were purchased from Cell Signaling. Rat Anti-HA (1:2,000) was purchased from Roche. Anti-HA agarose was from Pierce Biotechnology. Anti-GlyR was from Abcam (rabbit, 1:300) or Synaptic Systems (mouse, 1:300). Anti-PKC antibodies (sc-17804, A9, mouse, 1:500; sc-10800, H-300, rabbit 1:500) were from Santa Cruz Biotechnology.

Sun H., Lu L., Zuo Y., Wang Y., Jiao Y., Zeng W.Z., Huang C., Zhu M.X., Zamponi G.W., Zhou T., Xu T.L., Cheng J, & Li Y. (2014). Kainate receptor activation induces glycine receptor endocytosis through PKC deSUMOylation. Nature Communications, 5, 4980.

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SARS-CoV-2 RIP Assays in H1299-ACE2 Cells

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RIP assays were performed using the previously described method (Cao et al., 2018 (link); Keene et al., 2006 (link)). Briefly, H1299-ACE2 cells were transfected with pHA-tag, pHA-RBM24, pHA-ΔRNP1/2, pHA-ΔRNP1, or pHA-ΔRNP2, and infected with SARS-CoV-2 at MOI of 0.1, following which the cell lysates were harvested at 48 hpi. Then, 800 μg of the cell lysates containing the total protein was incubated with protein G precoated with mouse anti-HA (H9658; Sigma) at 30 °C for 4 h. After washing 5 times with washing buffer, the precipitated RNA and 5% of the total RNA (input RNA) extracted from the cell lysates were detected by real-time PCR. The relative level of the precipitated RNA of SARS-CoV-2 was normalized to that of the input RNA.

Yao Y., Sun H., Chen Y., Tian L., Huang D., Liu C., Zhou Y., Wang Y., Wen Z., Yang B., Chen X, & Pei R. (2022). RBM24 inhibits the translation of SARS-CoV-2 polyproteins by targeting the 5ʹ-untranslated region. Antiviral Research, 209, 105478.

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Molecular Reagents and Antibodies for Cell Signaling

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Tris, NaCl, and SDS for molecular biology and buffer preparation were purchased from Sigma‐Aldrich. Cell culture media and supplements were from Invitrogen (La Jolla, CA, USA). Antibodies were purchased from commercial sources as follows: anti‐EGFR (IHC‐00005; Bethyl Laboratories, Montigny, TX, USA), anti‐phospho‐EGFR [Tyr1068, Tyr992, and Tyr845 (Cell Signaling, Danvers, MA, USA)], anti‐TRAF4 (sc‐1920; Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse anti‐HA (H9658; Sigma‐Aldrich), Rabbit anti‐HA (Cell Signaling), mouse anti‐Omni and rabbit anti‐Omni (Santa Cruz Biotechnology), rabbit anti‐myc tag (Cell Signaling), mouse anti‐V5 antibody (Invitrogen), anti‐phospho‐ERK5 (Cell Signaling), anti‐ERK5 (Cell Signaling), anti‐phospho‐ERK1,2 (Santa Cruz Biotechnology), anti‐ERK1,2 (Cell Signaling), anti‐AKT (Cell Signaling), anti‐phospho‐AKT (Ser473; Cell Signaling), anti‐β‐actin (Cell Signaling). Recombinant human EGF was purchased from R&D (Minneapolis, MN, USA). Doxycycline and DMSO were purchased from Sigma‐Aldrich.

He S., Dong D., Lin J., Wu B., Nie X, & Cai G. (2022). Overexpression of TRAF4 promotes lung cancer growth and EGFR‐dependent phosphorylation of ERK5. FEBS Open Bio, 12(10), 1747-1760.

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Immunoblotting Technique for Protein Detection

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Immunoblotting was performed as described (Buchanan et al., 2019 (link)). The following antibodies were used for immunoblotting: mouse anti-HA (H9658, Sigma) at 1:2,000; rabbit anti-Ubc6 (gift from Thomas Sommer) (Walter et al., 2001 (link)) at 1:10,000; mouse anti-FLAG (F3165; Sigma) at 1:10,000; mouse anti-HA (16B12, BioLegend) at 1:1,000; mouse anti-MYC (9E10, Covance) at 1:10,000; rabbit anti-Doa10 at 1:2,000 (Kreft et al., 2006 (link)); rabbit anti-Cue1 at 1:2,000 (Cohen et al., 2015 (link)); mouse anti-PGK (459250, Thermo Fisher) at 1:20,000; rabbit anti-G6PDH (A9521, Sigma) at 1:20,000. Primary antibody incubations were followed with either peroxidase-coupled sheep anti-mouse or peroxidase-coupled goat anti-rabbit secondary antibodies (GE Healthcare) at 1:10,000 and visualized by enhanced chemiluminescence (Mruk and Cheng, 2011 (link)) with imaging on a G:Box system (Syngene).

Mehrtash A.B, & Hochstrasser M. (2022). Elements of the ERAD ubiquitin ligase Doa10 regulating sequential poly-ubiquitylation of its targets. iScience, 25(11), 105351.

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