Mouse anti ha 11 clone 16b12

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Mouse anti-HA.11 (clone 16B12) is an immunoglobulin G (IgG) monoclonal antibody that specifically binds to the influenza hemagglutinin (HA) epitope tag. This antibody is commonly used for the detection and purification of HA-tagged proteins in various applications.

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Anti-HA.11 (16 citations) Mouse anti-HA (clone 16B12; (8 citations) Mouse anti-HA (16B12, (5 citations) Mouse anti-HA.11 (4 citations) Anti-HA (16B12, (4 citations) Anti-HA.11 clone 16B12 (3 citations) Anti-HA (clone 16B12, (3 citations) Mouse HA.11 clone 16B12 (2 citations) Mouse anti-HA (HA.11) (2 citations) 16B12 Clone (1 citations)

8 protocols using mouse anti ha 11 clone 16b12

Monoclonal Anti-IAV NP Antibody Purification

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Doxycycline hyclate (Dox) was purchased from Sigma Aldrich. Hybridoma
cells secreting mouse monoclonal anti-IAV NP (clone H16-L10-4R5, ATCC
HB-65)^{25 (link)} were obtained
from ATCC and antibodies in the supernatant purified using a protein G column.
Polyclonal rabbit anti-neomycin phosphotransferase II (NPTII) was purchased from
Fitzgerald Industries International. Mouse anti-HA.11 (clone 16B12) was acquired
from BioLegend, polyclonal rabbit anti-HA (Y-11) from Santa Cruz. Mouse
anti-HA.11 (clone 16B12) coupled to Alexa Fluor (AF) 488 or AF594, as well as
fluorescently-labeled secondary antibodies and AF647 Phalloidin were obtained
from Life Technologies.

Schmidt F.I., Hanke L., Morin B., Brewer R., Brusic V., Whelan S.P, & Ploegh H.L. (2016). Phenotypic lentivirus screens to identify functional single domain antibodies. Nature microbiology, 1(8), 16080.

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Purification and Labeling of Antibodies

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Doxycycline hyclate (Dox) was purchased from Sigma-Aldrich. Nickel-nitrilotriacetic acid (NTA) beads were purchased from Qiagen. Mouse anti-HA.11 (clone 16B12) coupled to Alexa Fluor (AF) 488 was purchased from Life Technologies. Mouse anti-HA.11 (clone 16B12) was acquired from BioLegend. VHHs coupled to either Alexa Fluor (AF) 647 or 5-carboxytetramethylrhodamine (TAMRA) were generated using sortase A as described earlier (44 (link)). Hybridoma cells secreting mouse monoclonal anti-IAV NP (clone H16-L10-4R5; ATCC HB-65) were obtained from ATCC, and antibodies in the supernatant were purified using a protein G column.

Hanke L., Knockenhauer K.E., Brewer R.C., van Diest E., Schmidt F.I., Schwartz T.U, & Ploegh H.L. (2016). The Antiviral Mechanism of an Influenza A Virus Nucleoprotein-Specific Single-Domain Antibody Fragment. mBio, 7(6), e01569-16.

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Antibody Characterization for LC3B and p62

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Antibodies and their manufacturers were: mouse anti-FLAG clone M2 (WB 1 : 4000, IF 1 : 400; F1804) and rabbit anti-LC3B (WB 1 : 1000, IF 1 : 200; L7543) were from Sigma–Aldrich; goat anti-FLAG (1 : 400; ab1257), rabbit anti-SQSTM1/p62 (IP 1–2 μg/mg of lysate; ab101266) and rabbit anti-GFP (WB 1 : 2000, ab290) were from Abcam; mouse anti-SQSTM1/p62 (WB 1 : 1000 IF 1 : 200; 610832) was from BD Biosciences; rabbit anti-SQSTM1/p62 (WB 1 : 1000, 8025S) was from Cell Signaling Technology; mouse anti-HA.11 clone 16B12 (WB 1 : 1000, 901501) was from Biolegend; mouse anti-HA clone 12CA5 (WB 1 : 2000; 11583816001) from Roche; mouse IgG isotype control (14-4732-82) was from Invitrogen; Alexa Fluor 488-, 555-, 594- and 647-conjugated secondary antibodies were from Thermo Fisher (A21206, A31570, A11032 and A21447, respectively).

Ylä-Anttila P, & Masucci M.G. (2021). Inhibition of selective autophagy by members of the herpesvirus ubiquitin-deconjugase family. Biochemical Journal, 478(12), 2297-2308.

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Reagents and Antibodies for Inflammasome Assays

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Doxycycline hyclate was purchased from Sigma-Aldrich; E. coli K12 LPS, nigericin, and poly(dA:dT) were obtained from InvivoGen. PMA was purchased from Santa Cruz Biotechnology, Inc.; TAK-242 was obtained from EMD Millipore; and z-YVAD-cmk was purchased from Bio-Techne. Mouse anti-HA.11 (clone 16B12) was acquired from BioLegend, mouse anti-EGFP (clone JL-8) was purchased from Takara Bio Inc., mouse anti–IL-1β (clone 3A6) was obtained from Cell Signaling Technology, rabbit anti–caspase-1 p10 (C-20) was purchased from Santa Cruz Biotechnology, Inc., and rabbit anti-ASC (AL177) was obtained from Adipogen (all antibodies against the human homologues). HRP-coupled sheep anti–mouse IgG was purchased from GE Healthcare, HRP-coupled goat anti–rabbit IgG was obtained from SouthernBiotech, HRP-coupled goat anti–llama IgG was obtained from Bethyl Laboratories, peroxidase-coupled rat anti-HA (clone 3F10) was obtained from Sigma Aldrich, and HRP-coupled mouse anti-GAPDH (clone 6C5) was purchased from Abcam. Fluorescent secondary antibodies were acquired from Life Technologies.

Schmidt F.I., Lu A., Chen J.W., Ruan J., Tang C., Wu H, & Ploegh H.L. (2016). A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly. The Journal of Experimental Medicine, 213(5), 771-790.

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Immunofluorescence Antibody Detection Protocol

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Mouse anti-HA.11 (clone 16B12) and mouse anti-His tag (clone J099B12) were purchased from BioLegend. Rat anti-HA (clone 3F10) was purchased from Roche. Rabbit anti-TgSAG1 antibodies was provided by John Boothroyd (Stanford University), rat anti-TgSAG1 was provided by Vernon Carruthers (University of Michigan), and rabbit anti-TgGRA7 was provided by David Sibley (Washington University in St. Louis). Goat secondary antibodies conjugated to infrared (IR) dyes and Alexa Fluor dyes were purchased from Li-Cor and Invitrogen, respectively.

Moss W.J., Patterson C.E., Jochmans A.K, & Brown K.M. (2022). Functional Analysis of the Expanded Phosphodiesterase Gene Family in Toxoplasma gondii Tachyzoites. mSphere, 7(1), e00793-21.

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Antibody Characterization for Protein Studies

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Antibodies and their manufacturers were: mouse anti-FLAG clone M2 (1:5000, IF: 1:500; F1804) from Sigma-Aldrich; mouse anti-HA clone 12CA5 (1:2000; 11583816001) from Roche; mouse anti-pan 14-3-3 clone H-8 (1:1000; sc-1657), from Santa-Cruz Biotechnology (Santa Cruz, CA, USA); rabbit anti-IRF-3 clone D6I4C (1:1000, IF 1:200; #11904) and mouse anti-GST clone 26H1 (1:1000, IF 1:100 #2624) from Cell-Signaling Technologies (Danvers, MA, USA); rabbit anti TRIM25 clone EPR7315 (1:2000, IF: 1:100; ab167154) from Abcam (Cambridge, MA, USA); mouse anti-HA.11 clone 16B12 (1:1000; 901501) from BioLegend (San Diego, CA, USA). Alexa Fluor 488, 555, and 647 conjugated secondary antibodies were from Thermo Fisher (A21206, A31570, and A21447, respectively).

Gupta S., Ylä-Anttila P., Sandalova T., Achour A, & Masucci M.G. (2020). Interaction With 14-3-3 Correlates With Inactivation of the RIG-I Signalosome by Herpesvirus Ubiquitin Deconjugases. Frontiers in Immunology, 11, 437.

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Investigating EDEM3 and Protein Trafficking

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HEK293T and HeLa cells were from the European Collection of Animal Cell Cultures. Rabbit anti-EDEM3 (E8906) and rat anti-HA, clone 3F10 (cat: 12158167001) were from Sigma-Aldrich (St. Louis, MO, USA). Mouse anti-HA.11, clone 16B12 (cat: 901503) was from BioLegend (San Diego, CA, USA). Rabbit anti-α1-anti-trypsin (A0012) was from Dako (Jena, Germany). Mouse anti-tyrosinase (T311) antibody (sc-20035) and mouse anti-MAN1B1 antibody (sc-393145) was from Santa Cruz Biotechnology (Dallas, TX, USA). Rabbit polyclonal antibodies were generated in house for tyrosinase [63 ] and EDEM3 [64 ]. Rabbit anti-calnexin (ab22595), rat anti-tubulin (ab6161) and mouse anti-actin (ab8224) antibodies were from Abcam (Cambridge, UK). Endoglycosidase H (EndoH, P0702S) and Peptide-N-Glycosidase F (PNG-ase F, P0704S) were from New England Bioland (Ipswich, MA, USA). Kifunensine (sc-201364), MG132 (sc-201260) and all other chemicals were from Santa Cruz Biotechnology unless specified.

Manica G., Ghenea S., Munteanu C.V., Martin E.C., Butnaru C., Surleac M., Chiritoiu G.N., Alexandru P.R., Petrescu A.J, & Petrescu S.M. (2021). EDEM3 Domains Cooperate to Perform Its Overall Cell Functioning. International Journal of Molecular Sciences, 22(4), 2172.

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Monoclonal Anti-IAV NP Antibody Purification

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