HEK293, HEK293Trex, and MEF cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM) with 10% fetal bovine serum (FBS). Cycloheximide (CHX), and puromycin were purchased from Sigma. Anti-RPS6 (cat#2217), anti-phospho-eIF2α (cat#3398), anti- eIF2α (cat#5324), anti-phospho-4EBP1 (cat#9459), and anti-4EBP1 (cat#9452) antibodies were purchased from Cell Signaling. Anti-RPL5 antibody (SAB1100578) was from Sigma. Anti-HA antibody (SC-7392) was from Santa Cruz Biotechnology. Plasmids transfection was performed using Lipofectamine 2000 (Invitrogen) following the manufacturer's instruction.
Anti ha antibody sc 7392
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-HA antibody (sc-7392) is a monoclonal antibody that recognizes the hemagglutinin (HA) tag, a commonly used epitope tag. This antibody can be used to detect and immunoprecipitate HA-tagged proteins in various applications such as Western blotting, immunoprecipitation, and immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Anti rps6, by Cell Signaling Technology (2 mentions)
Anti 4e bp1, by Cell Signaling Technology (2 mentions)
Anti phospho eif2α, by Cell Signaling Technology (2 mentions)
Anti phospho 4ebp1, by Cell Signaling Technology (2 mentions)
Anti rpl5 antibody sab1100578, by Merck Group (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Anti eif2α, by Cell Signaling Technology (2 mentions)
Puromycin, by Merck Group (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Pb210pa 1, by System Biosciences (2 mentions)
Ab8580, by Abcam (1 mentions)
Fugene hd, by Promega (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
8 protocols using anti ha antibody sc 7392
1
Protein Expression Analysis in Cell Lines
2
Protein Expression Analysis in Cell Lines
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HEK293, HEK293Trex, and MEF cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM) with 10% fetal bovine serum (FBS). Cycloheximide (CHX), and puromycin were purchased from Sigma. Anti-RPS6 (cat#2217), anti-phospho-eIF2α (cat#3398), anti- eIF2α (cat#5324), anti-phospho-4EBP1 (cat#9459), and anti-4EBP1 (cat#9452) antibodies were purchased from Cell Signaling. Anti-RPL5 antibody (SAB1100578) was from Sigma. Anti-HA antibody (SC-7392) was from Santa Cruz Biotechnology. Plasmids transfection was performed using Lipofectamine 2000 (Invitrogen) following the manufacturer's instruction.
3
Innate Immune Signaling Pathway Analysis
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Anti-p-IRF3 antibody (4947) and anti-TBK1 antibody (3504) were purchased from Cell Signaling; anti-IRF3 antibody (ab68481) was from abcam; ISD was synthesized from Invitrogen; cGAMP (tlrl-cga23) was purchased from InvivoGen; anti-HA antibody (sc-7392) was purchased from Santa Cruz Biotechnology; anti-Flag M2 affinity gel (a2220), PMA (phorbol 12-myristate 13-acetate; 524400) and HT-DNA (D6898) were purchased from Sigma-Aldrich. Anti-cGAS antibody was prepared in our laboratory.
4
Immunoprecipitation of HA-tagged Proteins
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Pierce™ protein A/G Magnetic Beads were incubated with anti-HA antibody (sc-7392, Santa Cruz Biotechnology) in 5% (w/w) BSA for 3 h at 4°C, using 10 µl beads and 1 µg antibody per IP. Beads were washed three times with IP lysis buffer (500 µl) and incubated overnight using an end-over-end rotor at 4°C. Beads were then collected using a magnetic stand and washed three times with wash buffer prior to elution of bound protein in 30 µl of 2× SDS loading buffer for SDS–PAGE and immunoblotting.
5
Transient Expression of Cpf1 Proteins in Soybean Protoplasts
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The plant-codon-optimized pAsCpf1 and pLbCpf1, and E. coli-codon-optimized eAsCpf1 and eLbCpf1 were chemically synthesized (Bioneer, South Korea), and the full sequences of those genes were confirmed by Sanger sequencing. The p2GW7 destination vector was used to transiently express Cpf1 proteins in protoplasts. All plasmid sequences are available in Supplementary Note 1 and accompanied by detailed descriptions.
To assess plasmid-based expression of Cpf1 in plant cells, we applied Cpf1-harbouring plasmids (20 μg) into soybean protoplasts via PEG-mediated transformation. The transformed protoplasts were harvested after 24 h incubation and applied to western blot analysis with anti-HA antibody (sc-7392; Santa Cruz Biotechnology; 1:200) for detecting Cpf1 and anti-Histone-H3 (tri methyl K4) antibody (ab8580; Abcam; 1 μg ml−1) for measuring amounts of loading proteins.
To assess plasmid-based expression of Cpf1 in plant cells, we applied Cpf1-harbouring plasmids (20 μg) into soybean protoplasts via PEG-mediated transformation. The transformed protoplasts were harvested after 24 h incubation and applied to western blot analysis with anti-HA antibody (sc-7392; Santa Cruz Biotechnology; 1:200) for detecting Cpf1 and anti-Histone-H3 (tri methyl K4) antibody (ab8580; Abcam; 1 μg ml−1) for measuring amounts of loading proteins.
6
Inducible CPAP-HA Expressing HeLa Cell Line
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To generate a polyclonal cell line stably expressing inducible CPAP fused with HA tag, we used PiggyBac Transposon/Transposase genetic modification system. A cargo plasmid pCE0934_Dox-PB-CPAP-WT-2X-HA (modified from Wang et al. [2014] (link)) contained an siRNA resistant CPAP cDNA (with silent mutations in sequence 5′-AGAATTAGCTCGAATAGAAGA3′ to 5′-AGAGTTAGCTAGGATCGAAGA3′; Kohlmaier et al., 2009 (link)), tagged with two HA sequences on its C terminus and was flanked with inverted terminal repeats. The cargo plasmid and SPB plasmid (PB210PA-1; System Biosciences) expressing transposase were cotransfected to HeLaCPAP-AID cell line at a 4:1 ratio using FuGENE-HD (E2311; Promega), following manufacturer’s instructions. 2 d later, 2 µg/ml of puromycin was added to the culture medium for 10 d until puromycin-resistant colonies emerged. CPAP-HA was induced by 1 µg/ml doxycycline, and the presence of CPAP-HA was confirmed using anti-HA antibody (sc-7392; Santa Cruz) and CPAP-Ct antibody.
7
Visualizing Cas12a Expression in HEK293T Cells
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HEK293T cells were seeded on a 24-well plate. After 18 h, the cells were transiently transfected with Cas12a-crRNA-mRNA ternary complex. Eight hours after transfection, the medium was removed, and the cells were washed with PBS. The cells were then fixed with 4% paraformaldehyde for 10 min at room temperature, permeabilized with 0.25% Triton X-100 for 5 min at room temperature, and blocked in 5% BSA for 1 h at room temperature. The blocked cells were incubated with anti-HA antibody (sc-7392, Santa Cruz Biotechnology) at 1:100 dilution at room temperature for 1 h. After washing three times with PBS, the samples were incubated with the Goat anti-Mouse IgG (H + L) secondary antibody-FITC (#31569, Invitrogen) in the dark for 1 h at room temperature. The samples were washed thrice with PBS and stained with DAPI to visualize the nuclei. After washing five times, the coverslip samples were placed on microscope slides and dried overnight. The samples were visualized and imaged with fluorescence microscopy (Olympus).
8
Yeast Meiotic Cell Protein Lysate Preparation
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Yeast cell protein lysates were prepared as previously described [16] . Meiotic cells were harvested and centrifuged at 5,000 rpm for 1 min, and the pellet was resuspended in 100 µl of distilled-water; 100 µl of 0.6 M NaOH was then added to a final concentration of 0.3 M NaOH. Samples were incubated at room temperature for 5 min, and then centrifuged at 5,000 rpm for 1 min. Each pellet was resuspended in Laemmli buffer and boiled at 95°C for 5 min. Lysates were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). Mcd1 was detected using an anti-HA antibody (sc-7392; Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted 1:2,000.
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