Rat anti ha

Manufactured by Roche

Sourced in United States, Germany, Switzerland, France

Rat anti-HA is a monoclonal antibody that recognizes the hemagglutinin (HA) epitope tag. The HA epitope tag is a commonly used protein tag that allows for the detection and purification of recombinant proteins expressed in various systems. The Rat anti-HA antibody can be used in various applications, such as Western blotting, immunoprecipitation, and immunocytochemistry, to detect and analyze HA-tagged proteins.

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Lab products found in correlation

Mouse anti flag, by Merck Group (23 mentions) Mouse anti gfp, by Roche (15 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (9 mentions) Rabbit anti flag, by Merck Group (8 mentions) Rabbit anti myc, by Cell Signaling Technology (6 mentions) Mouse anti flag m2, by Merck Group (6 mentions) Mouse anti v5, by Thermo Fisher Scientific (6 mentions) Alexa 488, by Thermo Fisher Scientific (6 mentions) Mouse anti α tubulin, by Merck Group (6 mentions) Lsm 880 confocal microscope, by Zeiss (6 mentions) Lsm 780 confocal microscope, by Zeiss (6 mentions) Vectashield, by Vector Laboratories (5 mentions) Mouse anti myc, by Merck Group (5 mentions) Hoechst, by Thermo Fisher Scientific (4 mentions) Mouse monoclonal anti gfp, by Roche (4 mentions) Mouse anti ha, by Fortrea (4 mentions) Rabbit anti ha, by Cell Signaling Technology (4 mentions) Trans blot turbo transfer system, by Bio-Rad (4 mentions) Rabbit anti ha, by Merck Group (4 mentions) Mouse anti myc, by Cell Signaling Technology (4 mentions)

169 protocols using rat anti ha

Immunofluorescence Localization and Western Blotting of Proteins

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The localization of proteins by immunofluorescence in fixed MEF cells was performed as described previously 10 (link). The following antibodies were used: rabbit anti-TOM20 (Santa Cruz Biotechnology, sc-11415, 1:200), Alexa Fluor 647 anti-rabbit (Abcam, ab150079, 1:1000), mouse anti-FLAG (Sigma, F1804, 1:1000), Alexa Fluor 594 anti-mouse (Life Technologies, R37121, 1:1000), rat anti-HA (Roche, 11867431001, 1:200), Alexa Fluor 488 anti-rat (Life Technologies, A11006). Immunofluorescence images were captured using a Zeiss LSM880 confocal microscope and processed using ImageJ.
Detection of proteins by western blotting was achieved by resolving 20-100 μg of extracted protein on SDS-PAGE 4-12% Bis-Tris Bolt gels. These were transferred to nitrocellulose using an iBlot 2 transfer cell (Life Technologies). Antibodies used for western blotting in this work: rat anti-HA (Roche, 11867431001, 1:500), goat anti-rat HRP (Santa Cruz, SC2065, 1:1000). Gels were stained for loading using Coomassie Brilliant Blue (Life Technologies).

Gammage P.A., Viscomi C., Simard M.L., Costa A.S., Gaude E., Powell C.A., Van Haute L., McCann B.J., Rebelo-Guiomar P., Cerutti R., Zhang L., Rebar E.J., Zeviani M., Frezza C., Stewart J.B, & Minczuk M. (2018). Genome editing in mitochondria corrects a pathogenic mtDNA mutation in vivo. Nature medicine, 24(11), 1691-1695.

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Immunofluorescence and Western Blot Analyses

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Localisation of proteins by immunofluorescence was carried out in fixed 143B cells as previously described (22 (link)). Images were captured using a Zeiss LSM 880 confocal microscope.
The following antibodies were used for immunofluorescence experiments in this work: mouse anti-TOM22 (Abcam, ab10436, 1:250), Alexafluor-594 anti-mouse (Molecular Probes, A11005, 1:1000), rabbit anti-TFAM (gifted by Prof. Rudolf Wiesner, 1:500), Alexafluor-405 anti-rabbit (Molecular Probes, A31553, 1:1000), rat anti-HA (Roche, 11867431001, 1:500), Alexafluor-488 anti-rat (Molecular Probes, A11006, 1:1000). Mounting medium used was either ProLong Gold Antifade Mountant (Molecular Probes), or ProLong Gold Antifade Mountant with DAPI (Molecular Probes).
For western blot analyses, ∼20 μg of extracted proteins were resolved on SDS-PAGE 4–12% bis-tris gels (Life Technologies). The following antibodies were used for western blotting in this work: mouse anti-FLAG (Sigma, F1804, 1:2000), rabbit anti-FLAG (Sigma, F7425, 1:2000), rat anti-HA (Roche, 11867431001, 1:1000), rabbit anti-Histone H4 (Abcam, ab10158, 1:5000), rabbit anti-SSB1 (kindly gifted by Prof D. Kang, 1:4000), mouse anti-TOM22 (Abcam, ab10436, 1:5000), mouse anti-GAPDH (Abcam, ab9484, 1:10 000), goat anti-rabbit HRP (Promega, W401B, 1:2000), goat anti-mouse HRP (Promega, W402B, 1:2000), goat anti-rat HRP (Santa Cruz, SC2065, 1:1000).

Gammage P.A., Gaude E., Van Haute L., Rebelo-Guiomar P., Jackson C.B., Rorbach J., Pekalski M.L., Robinson A.J., Charpentier M., Concordet J.P., Frezza C, & Minczuk M. (2016). Near-complete elimination of mutant mtDNA by iterative or dynamic dose-controlled treatment with mtZFNs. Nucleic Acids Research, 44(16), 7804-7816.

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Immunofluorescence Localization and Western Blotting of Proteins

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Immunofluorescence Assay for Plasmodium Proteins

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Primary anti-CPN60 (rabbit) antibodies [51 (link)] were used at a dilution of 1:3000, anti-IMC1 at 1:1000, anti-HA (Rat, Roche) at 1:500, and anti-ATRx1 (Mouse) at 1:1000. Secondary AlexaFluor 488- and 546-conjugated anti-rat and anti-rabbit antibodies (Life Technologies) were used at 1/10000 dilutions, respectively. Parasites were fixed in PBS containing 4% paraformaldehyde for 30 min on ice. Samples were permeabilized with 0.1% Triton X-100 in PBS for 10 min on ice before blocking in PBS containing 3% BSA and incubation with primary antibodies then secondary antibodies diluted in the blocking solution. Labelled parasites were stained with Hoechst (1:10,000, Life Technologies) for 20 min and then washed three times in PBS then H₂O. Coverslips were mounted onto slides prior to observation using a Zeiss epifluorescent microscope.

Amiar S., MacRae J.I., Callahan D.L., Dubois D., van Dooren G.G., Shears M.J., Cesbron-Delauw M.F., Maréchal E., McConville M.J., McFadden G.I., Yamaryo-Botté Y, & Botté C.Y. (2016). Apicoplast-Localized Lysophosphatidic Acid Precursor Assembly Is Required for Bulk Phospholipid Synthesis in Toxoplasma gondii and Relies on an Algal/Plant-Like Glycerol 3-Phosphate Acyltransferase. PLoS Pathogens, 12(8), e1005765.

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Antibody Acquisition and Usage

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All chemicals were obtained from Sigma-Aldrich. Primary antibodies were obtained
as follows; anti-FLAG (M2; Sigma) (mouse), anti-Myc (rabbit) (Cell Signaling),
and anti-HA (rat) (Roche Applied Science). All secondary antibodies were
obtained from Roche Applied Science.

Malik S., Jang W, & Kim C. (2017). Protein Interaction Mapping of Translational Regulators Affecting Expression of the Critical Stem Cell Factor Nos. Development & Reproduction, 21(4), 449-456.

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Immunoprecipitation Using Commercial Antibodies

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All chemicals were obtained from Sigma–Aldrich. Primary antibodies were obtained as follows; anti-FLAG (M2; Sigma) (mouse), anti-Myc (rabbit) (Cell Signaling), and anti-HA (rat) (Roche Applied Science). All secondary antibodies were obtained from Roche Applied Science.

Malik S., Jang W., Park S.Y., Kim J.Y., Kwon K.S, & Kim C. (2019). The target specificity of the RNA binding protein Pumilio is determined by distinct co-factors. Bioscience Reports, 39(6), BSR20190099.

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Dual-Labeling Immunohistochemistry in Mouse Brain

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After being deeply anesthetized with Ketamine and Xylazine, mice were perfused with phosphate buffer saline (PBS) and 4% paraformaldehyde in PBS. Brains were post-fixed overnight with 4%A PFA/PBS, embedded in 2% agarose /PBS, and then sectioned at 50 μm with a vibratome (Leica, Buffalo Grove, IL VT1000S). The following antibodies were used for immunohistochemistry: anti-GFP rabbit (Life Technologies, Thermo Fisher, Waltham, MA A-11122), anti-GFP chicken (Aves labs, Tigard, OR GFP-1020), anti-HA rat (Roche diagnostics, Indianapolis, IN clone 3F10). Whole slide images were taken with a microscope with 5x objective and XY-stage controlled by μManger (https://micro-manager.org). Grid/Collection stitching Fiji plugin (Preibisch et al., 2009 (link)) was used for image assembly. We followed the dual-color in situ protocol described in BraInSitu web site (http://www.nibb.ac.jp/brish/indexE.html) (Watakabe et al., 2006 (link)).

Shima Y., Sugino K., Hempel C.M., Shima M., Taneja P., Bullis J.B., Mehta S., Lois C, & Nelson S.B. (2016). A Mammalian enhancer trap resource for discovering and manipulating neuronal cell types. eLife, 5, e13503.

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Immunofluorescence Assay for Toxoplasma

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Primary antibodies anti-HA (Rat, Roche), anti-Inner Membrane Complex protein1 (IMC1), anti-SAG1, anti-proM2AP, and anti-AMA1 were used at dilutions 1:500, 1:1,000, 1:1,000, 1:1,000, and 1:1,000, respectively. Secondary Alexa Fluor 488- and 546-conjugated anti-mouse, anti-rat, and anti-rabbit antibodies (Life Technologies) were used at 1/2,500. For the IFA, parasites were grown on confluent HFF on coverslips and fixed in PBS containing 2.5% paraformaldehyde (PFA) for 15 min at room temperature (RT). Samples were permeabilized with 0.25% Triton X-100 in PBS for 10 min at RT prior to blocking in PBS containing 3% Bovine Serum Albumine (BSA) and subsequent incubation with primary antibodies then secondary antibodies diluted in the blocking solution. Labeled parasites were stained with Hoechst (1/10,000, Life Technologies) for 20 min and then washed three times in PBS before final mounting of the coverslips on a glass slide using fluorogel. The fluorescence was visualized using fluorescence microscope (Axio Imager 2_apotome; ZEISS).

Charital S., Shunmugam S., Dass S., Alazzi A.M., Arnold C.S., Katris N.J., Duley S., Quansah N.A., Pierrel F., Govin J., Yamaryo-Botté Y, & Botté C.Y. (2024). The acyl-CoA synthetase TgACS1 allows neutral lipid metabolism and extracellular motility in Toxoplasma gondii through relocation via its peroxisomal targeting sequence (PTS) under low nutrient conditions. mBio, 15(4), e00427-24.

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Measuring JNK3α2 Binding to Arrestin-3

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Binding of JNK3α2 to AT1R-bound arrestin-3 was measured by co-immunoprecipitation. HEK-293A cells were co-transfected with HA-tagged AT1R-Rluc8, Flag-JNK3α2, and GRK2 with or without arrestin-3. 48 h after transfection, cells in 60-mm plates were lysed in 0.4 ml of lysis buffer (50 mM Tris, 2 mM EDTA, 250 mM NaCl, 10% glycerol, 0.5% Nonidet P-40, 20 mM NaF, 1 mM Na orthovanadate, 10 mM N-ethylmaleimide, 2 mM benzamidine, and 1 mM phenylmethylsulfonyl fluoride) for 30 min at 4°C. After centrifugation, supernatants were pre-cleared by 30 μl of protein G agarose. Then 700 μl of lysates (combined from two 60 mm plates) were incubated with anti-HA (rat, Roche) antibody overnight followed by the addition of 30 μl of protein G agarose. The suspensions were transferred to centrifuge filters (Ultrafree, Millipore) and washed three times with 300 μl of lysis buffer. Proteins were eluted with 35 μl of SDS sample buffer and analyzed by Western blot.

Zhan X., Perez A., Gimenez L.E., Vishnivetskiy S.A, & Gurevich V.V. (2014). Arrestin-3 binds the MAP kinase JNK3α2 via multiple sites on both domains. Cellular signalling, 26(4), 766-776.

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Yeast Strain and Antibody Protocol

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Yeast (Saccharomyces cerevisiae) strains used in this paper are summarized in Table S1. Plasmids used in this study are summarized in Table S2 Antibodies used in this study included mouse monoclonal anti-GFP (Roche Diagnostics), rat anti-HA (Roche Diagnostics), mouse anti-HA (Covance), rabbit anti-G6PDH
(Sigma), rabbit anti-Ape1 (a kind gift from Dr. Ohsumi), rabbit anti-Atg8 (a kind gift from Dr.
Ohsumi), goat anti-rat HRP (Abcam), goat anti-rabbit HRP and goat anti-mouse HRP (GE Healthcare).

Lipatova Z., Gyurkovska V., Zhao S.F., & Segev N. (2020). Characterization of Constitutive macro-ER-phagy.

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