Rabbitmonoclonal anti ha antibody c29f4

Manufactured by Cell Signaling Technology

The Rabbit monoclonal anti-HA antibody (C29F4) is a laboratory reagent used for the detection and analysis of proteins tagged with the HA (Hemagglutinin) epitope. It is a highly specific antibody raised against the HA tag, which is a commonly used protein tag for various applications.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Polyethylenimine (pei), by Thermo Fisher Scientific (1 mentions) Ionomycin, by Thermo Fisher Scientific (1 mentions) Phenylmethanesulfonyl fluoride, by Merck Group (1 mentions) Anti erk1 2 antibody, by Santa Cruz Biotechnology (1 mentions) Hispur cobalt resin, by Thermo Fisher Scientific (1 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (1 mentions) Rneasy kit, by Qiagen (1 mentions) Restriction enzyme, by New England Biolabs (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions) Immobilon western, by Merck Group (1 mentions) Swine anti rabbit hrp, by Agilent Technologies (1 mentions) Goat anti mouse hrp, by Agilent Technologies (1 mentions) Mouse monoclonal anti tubulin t5168, by Merck Group (1 mentions) Blotting grade blocker, by Bio-Rad (1 mentions) Nucblue, by Thermo Fisher Scientific (1 mentions) Anti rabbit alexa488, by Thermo Fisher Scientific (1 mentions)

3 protocols using rabbitmonoclonal anti ha antibody c29f4

Protein Purification and Analysis Protocols

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RNeasy kit was purchased from Qiagen. High Capacity cDNA reverse
transcription kit was from Applied Biosystems (Foster City, CA). Restriction
enzymes and competent cells were purchased from New England Biolabs (Ipswich,
MA). PHUSION PCR reaction mix was from Thermo Fisher (Waltham, MA). Fetal bovine
serum and isopropyl β-D-1-thiogalactopyranoside were obtained from
Sigma-Aldrich (St. Louis, MO). HisPur cobalt resin was from Thermo Fisher
(Waltham, MA). Phenyl Sepharose CL-4B resin was from GE Healthcare Life Sciences
(Milwaukee, MI). DMEM culture medium was from Caisson Laboratories (Smithfield,
UT). Polyethylenimine and penicillin/streptomycin were from ThermoFisher
(Carlsbad, CA). Fura-2/AM was purchased from Teflabs (Austin, TX). Ionomycin and
XRhod-5F were obtained from ThermoFisher Scientific (Carlsbad, CA). Rabbit
monoclonal anti-HA antibody (C29F4) and anti-phospho-p44/42 MAPK mouse
monoclonal antibody (L34F12) were from Cell Signaling Technology (Danvers, MA).
Anti-ERK1/2 antibody was from Santa Cruz Biotechnology (SC514302, Santa Cruz,
CA). Protease inhibitor cocktail was from EMD Millipore (Burlington, MA).
Phenylmethane sulfonyl fluoride was from Sigma-Aldrich (St. Louis, MO). Amido
Black B was from Bio-Rad (Hercules, CA). A61603 and norepinephrine were from
Cayman Chemical (Ann Arbor, MI).

Gebert-Oberle B., Giles J., Clayton S, & Tran Q.K. (2019). Calcium/calmodulin regulates signaling at the α1A adrenoceptor. European journal of pharmacology, 848, 70-79.

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Western Blot Analysis of C. elegans Proteins

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Crowded plates of various C. elegans strains were harvested and washed twice in M9 and then lysed using the following lysis buffer: 25 mM Tris (pH 7.5), 300 mM NaCl, 1% triton X-100 and 1X protease inhibitor. 25 μg of total protein was loaded in each lane. For transgenic animals, 25 worms expressing GFP (used as a transformation marker) were picked, boiled in sample buffer (4x Laemmli sample buffer, BIO-RAD) and loaded on an SDS gel. For detection of protein the nitrocellulose membranes (GE Health care) were blocked with 5% skimmed milk (Blotting-Grade Blocker, BIO-RAD) diluted in PBS-T. Antibody dilutions (primary antibody: rabbit monoclonal anti-HA antibody (C29F4; Cell Signaling Technology) 1:5000, mouse monoclonal anti-TUBULIN (T5168; SIGMA) and secondary antibodies: swine anti-rabbit HRP (1:3000, Dako) or goat anti-mouse HRP (1:3000, Dako)) was done in 5% skimmed milk in TBST and washes were carried out in PBS-T. Detection of the bound antibodies was performed using an ECL detection kit (Immobilon Western; Millipore) and visualized with a digital camera (VersaDoc; Bio-Rad).

Busayavalasa K., Ruiz M., Devkota R., Ståhlman M., Bodhicharla R., Svensk E., Hermansson N.O., Borén J, & Pilon M. (2020). Leveraging a gain-of-function allele of Caenorhabditis elegans paqr-1 to elucidate membrane homeostasis by PAQR proteins. PLoS Genetics, 16(8), e1008975.

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Immunostaining of HEK293 cells

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HEK293 cells were fixed with 4% formaldehyde for 20 min at room temperature. Fixed cells were then washed with PBS and permeabilized with 0.1% saponin and blocked with 0.5% BSA and 0.5% gelatin for 60 min. Cells were washed once with washing buffer (0.01% saponin, 0.2% gelatin in PBS) and incubated with mouse monoclonal anti-FLAG M2 antibody (Sigma) and rabbit monoclonal anti-HA antibody (C29F4, Cell Signaling) for 60 min. Cells were then washed with washing buffer and incubated with secondary antibodies, anti-mouse Alexa 568 (Invitrogen) and anti-rabbit Alexa 488 (Invitrogen) for 60 min. After washing with washing buffer, mounting media with NucBlue (Invitrogen) was added to stain the nuclei. Images were acquired with an LSM880 confocal microscope with a 40× water-immersion objective. Pearson correlation coefficient was calculated with ImageJ, as described (23 ).

Ruiz M., Devkota R., Kaper D., Ruhanen H., Busayavalasa K., Radović U., Henricsson M., Käkelä R., Borén J, & Pilon M. (2023). AdipoR2 recruits protein interactors to promote fatty acid elongation and membrane fluidity. The Journal of Biological Chemistry, 299(6), 104799.

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