Biotinylated human IgG, goat-anti-chicken DyLight®633 (GAC633), goat-anti-chicken DyLight®487 (GAC487), donkey-anti-rabbit DyLight®633 (DAC633), goat-anti-human IgG DyLight®633, and goat-anti-chicken HRP conjugate were purchased from Immunoreagents (Raleigh, NC). Chicken anti-c-myc antibody, rabbit-anti-HA antibody, streptavidin R-phycoerythrin conjugate (SA-PE), Dynabeads™ Biotin Binder (streptavidin-coated beads), Dynabeads™ His-Tag Isolation and Pulldown beads, High-fidelity Phusion™ polymerase, and CloneJET™ PCR cloning kit (K1231) were purchased from Thermo-Fisher (Waltham, MA). Mouse anti-penta-His antibody-Alexa Fluor 647 conjugate and Ni-NTA agarose were obtained from Qiagen (Valencia, CA). Frozen-EZ Yeast Transformation Kit™, Zymoprep™ Yeast plasmid Miniprep II kit, Quick-DNA™ Miniprep Plus Kit were purchased from Zymo Research (Irvine, CA). All restriction enzymes were purchased from New England BioLabs (Ipswich, MA). Gene fragments were purchased from Integrated DNA Technologies (IDT; Coralville, IA). Oligonucleotide primers were purchased from IDT or Eton Biosciences (Raleigh, NC). Sequences of all primers and G-blocks are included in Table S1 (primers) and Table S2 (gene fragments). The composition of yeast culture media used was as previously described31 (link).
Rabbit anti ha antibody
Manufactured by Thermo Fisher Scientific
The Rabbit-anti-HA antibody is a primary antibody that specifically binds to the HA (Hemagglutinin) tag, a commonly used protein tag. It is produced in rabbits and can be used for the detection and identification of HA-tagged proteins in various applications such as Western blotting, immunoprecipitation, and immunofluorescence.
Automatically generated - may contain errors
Lab products found in correlation
Dynabeads his tag isolation and pulldown beads, by Thermo Fisher Scientific (1 mentions)
Chicken anti cmyc antibody, by Thermo Fisher Scientific (1 mentions)
Streptavidin r phycoerythrin conjugate, by Thermo Fisher Scientific (1 mentions)
Phusion high fidelity polymerase, by Thermo Fisher Scientific (1 mentions)
Clonejet pcr cloning kit, by Thermo Fisher Scientific (1 mentions)
Frozen ez yeast transformation kit, by Zymo Research (1 mentions)
Quick dna miniprep plus kit, by Zymo Research (1 mentions)
Zymoprep yeast plasmid miniprep 2 kit, by Zymo Research (1 mentions)
Ni nta agarose, by Qiagen (1 mentions)
Goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Sodium pyruvate s8636, by Merck Group (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Trypsin, by Merck Group (1 mentions)
Donkey serum, by Merck Group (1 mentions)
Alexa fluor 647 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions)
Sp8 confocal microscope, by Leica camera (1 mentions)
3 protocols using rabbit anti ha antibody
1
Protein Expression and Purification Protocol
2
Phosphorylation Analysis of Cellular Proteins
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Crystal violet (C‐3886), mouse anti‐phospho‐PKR (pThr446) antibody, anti‐c‐Myc (C3956) antibody, sodium pyruvate (S8636), and 0.25 mg/mL trypsin were purchased from Sigma‐Aldrich and used at the indicated final concentrations. Methylcellulose (118‐084‐721) was purchased from MP Biomedical. Anti‐eIF2α (9722), anti‐phospho‐eIF2α (Ser 51) (9721), and β‐actin antibodies were purchased from Cell Signaling Technology®. Mouse anti‐PKR (K‐17) antibody, goat anti‐mouse IgG, F(ab′)2‐HRP (sc‐3696), and goat anti‐rabbit IgG, F(ab′)2‐HRP (sc‐3837) secondary antibodies were purchased from Santa Cruz Biotechnology. Rabbit anti‐HA antibody was purchased from ThermoFisher Scientific. SPPV L1 protein antiserum was prepared by our laboratory.
3
Localization of PIEZO2 in N2A cells
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N2A cells were co-transfected with PIEZO2-HA-IRES-GFP or IDRdel constructs and with a plasmid encoding the red fluorescent protein mScarlet fused to a farnesylation signal sequence in its C-terminus to target it to the plasma membrane. Three days after transfection, cells were washed once with PBS and fixed with 4% PFA for 10 min at room temperature, washed 3 times for 5 min with PBS and permeabilised for 1 h at room temperature (permeabilization buffer: 2,5% donkey serum (Sigma), 1% BSA, 0.1% Triton X-100, 0.05% Tween-20, in PBS). Samples were then incubated overnight at 4 °C with a 1:500 dilution of rabbit anti-HA antibody (Thermo Fisher Scientific) in PBS 1% BSA. After 3 washes of 5 min, cells were incubated for 1 h with a 1:1000 dilution of AlexaFluor-647 donkey anti-rabbit (Life technologies, diluted in PBS 1% BSA) and washed 3 more times. Coverslips were mounted on slides with FluoProbe mounting media that contain DAPI (Interchim). Confocal images were acquired with a SP8 confocal microscope (Leica) and a 63x oil-immersion objective. Images were analyzed offline with Fiji (v2.3.0/1.53f).
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