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Ha tag antibody

Manufactured by Cell Signaling Technology
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The HA-tag antibody is a lab equipment product used for the detection and analysis of proteins tagged with the HA (Hemagglutinin) epitope. The HA-tag antibody specifically binds to the HA-tag sequence, allowing researchers to identify and track the expression of HA-tagged proteins in various experimental systems.

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Lab products found in correlation

Flag tag antibody, by Cell Signaling Technology (3 mentions) Normal rabbit mouse igg, by Cell Signaling Technology (3 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Pdd00017273, by Bio-Techne (1 mentions) Gfp antibody, by Thermo Fisher Scientific (1 mentions) Fugene hd transfection reagent, by Promega (1 mentions) Monoclonal anti flag m2 affinity gel, by Merck Group (1 mentions) Pdgfrb, by Cell Signaling Technology (1 mentions) Stat3, by Cell Signaling Technology (1 mentions) Phospho akt, by Cell Signaling Technology (1 mentions) β catenin antibody, by Cell Signaling Technology (1 mentions) Cy3 affinipure donkey anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Sp5 confocal microscope, by Leica (1 mentions) Lc3 antibody, by Merck Group (1 mentions) Bortezomib, by Merck Group (1 mentions) β tubulin, by Proteintech (1 mentions) Pepstatin a, by Merck Group (1 mentions) Cav 1, by Proteintech (1 mentions) Sclareol, by Merck Group (1 mentions) P erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

HA-tag (191 citations) Anti-HA-tag antibody (14 citations) Rabbit anti-HA tag antibody (7 citations) HA-Tag rabbit monoclonal antibody (4 citations) Mouse anti-HA-Tag monoclonal antibody (4 citations) HA-tag (C29F4) rabbit monoclonal antibody (3 citations) HA-Tag (6E2) mouse monoclonal antibody (3 citations) HA-Tag (6E2) mouse antibody (3 citations) Anti-HA tag antibody for HA-ins-PD-L1 (2 citations) HA-Tag Rabbit polyclonal antibody (2 citations)

12 protocols using ha tag antibody

1

Ubiquitination and ADP-Ribosylation Analysis

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COS-7 cells were co-transfected with a mixture of GFP, GFP-SET8 FL or GFP-SET8, 3xFLAG or 3xFLAG-PARP1, HA-ubiquitin plasmids, and Fugene HD transfection reagent (Promega, # E2311) according to manufacturer’s recommendations. After 48 h, transfected COS-7 cells were treated or not with 50 μM MG132 for 2 h and lysed as described previously60 (link). Synchronized GFP-SET8 FL transfected cells were treated for 3 h with 10 μM cullin inhibitor (MLN4924, Selleckem # S7109), PARG inhibitor (PDD 00017273, Tocris # 5952) or with DMSO as control. Cell lysates (100–200 μg) were then immunoprecipitated with GFP antibody (Thermo Fisher Scientific # G10362). Ubiquitination and ADP-ribosylation were detected by Western blot using HA-tag antibody (Cell signaling Technology # 3724S) or anti-ADPribose antibody, respectively.
Estève P.O., Sen S., Vishnu U.S., Ruse C., Chin H.G, & Pradhan S. (2022). Poly ADP-ribosylation of SET8 leads to aberrant H4K20 methylation in mammalian nuclear genome. Communications Biology, 5, 1292.
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2

Protein Interaction and Signaling Analysis Techniques

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Western blot, ELISA, and co-immunoprecipitation (Co-IP) were performed as previously described [18 (link)]. For Western blot analysis, protein samples were resolved on an SDS-PAGE gel and transferred to nitrocellulose membrane, followed by incubation with a primary antibody and an HRP-conjugated secondary antibody. Antibodies used in this work were the followings: Sesn3 (Rabbit pAb, PIPA522220, Fisher Scientific), Actinin (Mouse mAb, sc-17829, Santa Cruz Biotechnology), Pdgfrb (Rabbit mAb, #3169S, Cell Signaling Technology), phospho-Stat3 (Tyr705) (Rabbit mAb, #9145S, Cell Signaling Technology), Stat3 (Mouse mAb, #9139S, Cell Signaling Technology), phospho-Akt (Thr308) (Rabbit mAb, #4056S, Cell Signaling Technology), phospho-Akt (Ser473) (Rabbit mAb, #4058S, Cell Signaling Technology), and Akt (Rabbit mAb, #4685S, Cell Signaling Technology), To measure hepatic levels of IL-6, we used a commercial ELISA kit for mouse IL-6 (BD Biosciences) following the manufacturer’s instruction. Co-IP was performed in HEK 293 cells after co-transfection of indicated DNA constructs. IP and immunoblots were performed using tag antibodies: monoclonal anti-FLAG M2 affinity gel (Sigma) and HA tag antibody (Rabbit mAb, #3724S, Cell Signaling Technology).
Liu Y., Kim H.G., Dong E., Dong C., Huang M., Liu Y., Liangpunsakul S, & Dong X.C. (2019). Sesn3 deficiency promotes carcinogen-induced hepatocellular carcinoma via regulation of the hedgehog pathway. Biochimica et biophysica acta. Molecular basis of disease, 1865(10), 2685-2693.
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3

Protein-Protein Interaction Analysis

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Co-immunoprecipitation was performed as described previously [45 (link)]. Both the input and IP samples were analyzed by Western blot using various antibodies at the following dilutions: WISP1 antibody (1:1000), β-catenin antibody (1:1000), Flag-tag antibody (1:1000), HA-tag antibody (1:1000) and normal rabbit/mouse IgG (Cell Signaling Technology).
Wu J., Long Z., Cai H., Du C., Liu X., Yu S, & Wang Y. (2016). High expression of WISP1 in colon cancer is associated with apoptosis, invasion and poor prognosis. Oncotarget, 7(31), 49834-49847.
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4

Immunostaining of HA-tagged Proteins in HEK293 Cells

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Following KIFBP overexpression in HEK293, cells were fixed with 4% paraformaldehyde for 15 min, and made permeable with 1% bovine serum albumin and 0.1% Triton X‐100 in phosphate‐buffered saline (PBS). Cells were stained for HA using the HA‐Tag antibody (C29F4, Cell Signaling Technology, USA) at 1:1500 dilution, and the Cy3 AffiniPure donkey antirabbit IgG, 1:200 dilution (Jackson Immunoresearch, UK). Three wells were fixed for each construct. Cells were imaged on a Leica SP5 confocal microscope.
MacKenzie K.C., de Graaf B.M., Syrimis A., Zhao Y., Brosens E., Mancini G.M., Schot R., Halley D., Wilke M., Vøllo A., Flinter F., Green A., Mansour S., Pilch J., Stark Z., Zamba‐Papanicolaou E., Christophidou‐Anastasiadou V., Hofstra R.M., Jongbloed J.D., Nicolaou N., Tanteles G.A., Brooks A.S, & Alves M.M. (2020). Goldberg–Shprintzen syndrome is determined by the absence, or reduced expression levels, of KIFBP. Human Mutation, 41(11), 1906-1917.
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5

Antibody Procurement for Cellular Analyses

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Sclareol, bortezomib, E64 and pepstatin A were obtained from Sigma-Aldrich; Thermo Fisher Scientific, Inc. (Waltham, MA, USA). The Cav1, SOD1, β-tubulin and p62 antibodies were obtained from ProteinTech Group, Inc., Chicago, IL, USA), the LC3 antibody was obtained from Sigma-Aldrich; Thermo Fisher Scientific, Inc.), the Flag antibody was obtained from Prospec-Tany TechnoGene, Ltd., (East Brunswick, NJ, USA), the HA tag antibody was obtained from Cell Signaling Technology, Inc. (Beverly, MA, USA).
Zhang T., Wang T, & Cai P. (2017). Sclareol inhibits cell proliferation and sensitizes cells to the antiproliferative effect of bortezomib via upregulating the tumor suppressor caveolin-1 in cervical cancer cells. Molecular Medicine Reports, 15(6), 3566-3574.
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6

Antibody Use for Cell Signaling

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The BCKDK (E-12) (1:1000, sc-374425) antibodies were purchased from Santa Cruz Technology. Flag tag and talin-1 (#4021) antibodies were purchased from Sigma-Aldrich. All the following antibodies were purchased from Cell Signaling Technology: ERK1/2 (#4695), p-ERK1/2 (Thr202/Tyr204) (#4649), MEK1/2 (8727s), p-MEK1/2 (S221) (2338s), β1-integrin (D6S1W), TRIM21 (#92043), AKT (#4060), p-AKT (S473) (#13038), HA tag antibody (#3724), β-actin mouse monoclonal antibody (#3700), Anti-rabbit IgG (Alexa Fluor#488 Conjugate) (4412), Anti-mouse IgG (Alexa Fluor#594 Conjugate) (8890), secondary antibody against mouse and rabbit. The FAK (A11131), pY397-FAK (AP0302), talin-2 (A19810), and BCAT2 (A7426) antibodies were purchased from ABclonal. The vinculin (ab129002), BCKDHA (ab305168), p-BCKDHA (S293) (ab302504) antibodies were purchased from Abcam. All antibodies were used following the manufacturer’s instructions.
Xu C., Yang K., Xuan Z., Li J., Liu Y., Zhao Y., Zheng Z., Bai Y., Shi Z., Shao C., Zhang L, & Sun H. (2023). BCKDK regulates breast cancer cell adhesion and tumor metastasis by inhibiting TRIM21 ubiquitinate talin1. Cell Death & Disease, 14(7), 445.
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7

Immunofluorescence Staining Protocol

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Cells were washed twice with PBS, fixed with 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 in PBS for 15 min, and blocked with normal goat serum (Boster) for 1 hr at room temperature. Following, cells were incubated with HA tag antibody (1:500, Cell Signaling Technology) or pan-keratin antibody (1:500, Cell Signaling Technology). After 2 hr incubation, cells were washed for four times (5 min per washing) with PBST on table concentrator. Then secondary antibody conjugated 488 or 594 nm fluorophore (ThermoFisher) was incubated for 1 hr, and nuclei were stained with Hoechst 33342 (ThermoFisher). Images were captured with Leica confocal microscope.
Huang R., Fu Y, & Deng Y. (2021). KLF4 transactivates TRIM29 expression and modulates keratin network. Biochemistry and Biophysics Reports, 28, 101117.
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8

Co-Immunoprecipitation of GSK3β and Pit-1

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HEK293T cells (293T) were seeded overnight at a density of 10 × 106/per 15-cm plate in DMEM supplemented with 10% fetal bovine serum. Cells were cotransfected with a human GSK3βWT-HA–tagged plasmid (14753, Addgene; provided by Dr Jim Woodgett, University of Toronto, Toronto, Ontario, Canada) together with either pcDNA3.1-Myc/His (Invitrogen)– or human Pit-1-Myc–tagged plasmid. At 36 hours posttransfection, nuclear lysates were collected, and coimmunoprecipitation was performed with the Universal Magnetic Co-IP Kit with 500 μg of nuclear lysate per condition either with 2 μg of Myc-Tag Antibody (2276; Cell Signaling Technology) or with the IgG isotype control (Cell Signaling Technology). Samples were prepared as mentioned previously, and the interaction between GSK3-β and Pit-1 was determined by sequential blotting with HA-tag antibody (3724, 1:1000; Cell Signaling) and Myc-Tag Antibody (2276, 1:1000; Cell Signaling Technology). Input protein equal to 1% of protein used in the immunoprecipitation reactions was loaded as a control in a separate gel similarly blotted with HA- and Myc-tag antibodies.
Eigler T., Ben-Shlomo A., Zhou C., Khalafi R., Ren S.G, & Melmed S. (2014). Constitutive Somatostatin Receptor Subtype-3 Signaling Suppresses Growth Hormone Synthesis. Molecular Endocrinology, 28(4), 554-564.
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9

Co-immunoprecipitation and Western Blot Analysis

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Co-immunoprecipitation was performed as described previously [30 (link)]. Both the input and IP samples were analyzed by Western blot using various antibodies at the following dilutions: TRIM65 antibody (1:1000), RhoA antibody (1:1000), Flag-tag antibody (1:1000), HA-tag antibody (1:1000) and normal rabbit/mouse IgG (Cell Signaling Technology)
Wang X.L., Shi W.P., Shi H.C., Lu S.C., Wang K., Sun C., He J.S., Jin W.G., Lv X.X., Zou H, & Shu Y.S. (2016). Knockdown of TRIM65 inhibits lung cancer cell proliferation, migration and invasion: A therapeutic target in human lung cancer. Oncotarget, 7(49), 81527-81540.
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10

Co-immunoprecipitation of FKBP14 and RhoA

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Co-immunoprecipitation was performed as described previously [45 (link)]. Both the input and IP samples were analyzed by Western blot using various antibodies at the following dilutions: FKBP14 antibody (1:1000), RhoA antibody (1:1000), Flag-tag antibody (1:1000), HA-tag antibody (1:1000) and normal rabbit/mouse IgG (Cell Signaling Technology).
Huang Z., Li J., Du S., Tang Y., Huang L., Xiao L, & Tong P. (2016). FKBP14 overexpression contributes to osteosarcoma carcinogenesis and indicates poor survival outcome. Oncotarget, 7(26), 39872-39884.
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