Mouse monoclonal anti ha

Manufactured by BioLegend

The Mouse monoclonal anti-HA is a laboratory reagent used for the detection and identification of HA-tagged proteins. It is a mouse-derived monoclonal antibody that specifically binds to the HA epitope tag, which is commonly used to facilitate the purification and detection of recombinant proteins.

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Lab products found in correlation

Rabbit anti actin, by Merck Group (2 mentions) Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions) Lysotracker, by Thermo Fisher Scientific (1 mentions) Mouse anti tubulin, by Merck Group (1 mentions) Mouse anti rab7, by Santa Cruz Biotechnology (1 mentions) Mouse monoclonal anti lamp2, by Abcam (1 mentions) Mouse anti rack1, by BD (1 mentions) Mouse anti eea1, by Santa Cruz Biotechnology (1 mentions) Control short hairpin rna shrna, by Merck Group (1 mentions) Nupage novex bis tris gel, by Thermo Fisher Scientific (1 mentions) Mouse monoclonal anti flag clone m2, by Merck Group (1 mentions) Anti znf750, by Merck Group (1 mentions) Rabbit polyclonal anti flag, by Merck Group (1 mentions) Mouse monoclonal anti vinculin, by Merck Group (1 mentions) Anti krt10, by Fortrea (1 mentions) Anti loricrin, by Fortrea (1 mentions) Rabbit monoclonal anti ha, by Cell Signaling Technology (1 mentions) Mouse monoclonal anti gfp, by Roche (1 mentions) Anti tap cab1001, by Thermo Fisher Scientific (1 mentions) Goat anti rabbit igg h l hrp conjugate, by Bio-Rad (1 mentions)

Spelling variants of this product

Anti-HA (86 citations) Mouse anti-HA (41 citations) Mouse monoclonal anti-HA.11 (ascites, (2 citations) Mouse monoclonal anti-hemagglutinin (HA) (2 citations) Mouse monoclonal anti-HA.11 (2 citations) Mouse monoclonal anti-HA antibody (2 citations) Monoclonal mouse anti-HA.11 antibody (2 citations)

7 protocols using mouse monoclonal anti ha

Subcellular Localization and Trafficking Analysis

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Antibodies used for immunofluorescence microscopy or western blotting were obtained from commercial sources. The following antibodies were used: rabbit anti-NHE6 (Abcam; 137185), rabbit anti-NHE9 (Abcam; 167157), mouse anti-RACK1 (BD Biosciences; 610177), rabbit anti-RACK1 (Cell Signaling Technology; 5432), mouse anti-tubulin (Sigma-Aldrich; T6199), rabbit anti-actin (Sigma-Aldrich; A5060), mouse monoclonal anti-HA (BioLegend; MMS-101P), mouse anti-EEA1(Santa Cruz Biotechnology; 6415), mouse anti-Rab7 (Santa Cruz Biotechnology; 10767), rabbit phospho-(Ser) PKC substrate (Cell Signaling Technology; 2261), mouse monoclonal PKC (Santa Cruz Biotechnology; 80), mouse monoclonal anti-LAMP2 (Abcam; 25631), rabbit anti-p-glycoprotein (Abcam; 129450). All Alexa Fluor secondary antibodies were acquired from Thermo Fisher Scientific. 4′, 6-diamidino-2-phenylindol dilactate (DAPI), Alexa Fluor-conjugated Tfn and Lysotracker were purchased from Thermo Fisher Scientific. Chemotherapeutic drugs (Dox, Mtx, Dau) were obtained from the local drug dispensary of the Centre Hospitalier Universitaire de Sherbrooke. Cq and Baf were obtained from Sigma-Aldrich. Cq was dissolved in water and Baf in dimethylsulfoxide. Control short hairpin RNA (shRNA) and shRNA directed against human NHE6 or NHE9 were purchased from Sigma-Aldrich. RNA interference directed against RACK1 was purchased from Ambion.

Lucien F., Pelletier P.P., Lavoie R.R., Lacroix J.M., Roy S., Parent J.L., Arsenault D., Harper K, & Dubois C.M. (2017). Hypoxia-induced mobilization of NHE6 to the plasma membrane triggers endosome hyperacidification and chemoresistance. Nature Communications, 8, 15884.

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Western Blot Analysis of Protein Targets

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30~50μg total protein per lane was separated on 10% Novex NuPAGE Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membrane following standard procedures. The following antibodies are used for western blotting: mouse monoclonal anti-GFP (sc-9996), mouse monoclonal anti-HA (Biolegend, 901501), mouse monoclonal anti-Araf (sc-166771) and custom rabbit anti-serum (Covance) that is able to recognize both endogenous mouse PABP and conditionally expressed PABP-GFP.

Hwang H.W., Saito Y., Park C.Y., Blachère N.E., Tajima Y., Fak J.J., Zucker-Scharff I, & Darnell R.B. (2017). cTag-PAPERCLIP reveals alternative polyadenylation promotes cell-type specific protein diversity and switches Araf isoforms with microglia activation. Neuron, 95(6), 1334-1349.e5.

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Diverse Antibody Utilization in Research

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We utilized the following antibodies: mouse monoclonal anti-FLAG (clone M2, Sigma-Aldrich), rabbit polyclonal anti-FLAG (Sigma-Aldrich), rabbit polyclonal anti-Senataxin (Novus Biologicals, NBP1-94712), rabbit polyclonal anti-Senataxin (kind gift from Domenico Delia, University of Milan, Milan, Italy), mouse monoclonal anti-HA (BioLegend), rabbit monoclonal anti-p63-α (clone D2K8X; Cell Signaling Technology), mouse monoclonal anti-DNA–RNA hybrids clone S9.6 (Millipore), mouse monoclonal anti-vinculin (Sigma), rabbit polyclonal anti‐KRT1 (BioLegend, PRB‐149P), rabbit polyclonal anti‐KRT10 (Covance PRB159P), rabbit polyclonal anti ZNF750 (Sigma-Aldrich, HPA023012), rabbit polyclonal anti-Loricrin (Covance PRB145P), and rabbit polyclonal anti-Adenosine (Biovision).

Gatti V., Fierro C., Compagnone M., La Banca V., Mauriello A., Montanaro M., Scalera S., De Nicola F., Candi E., Ricci F., Fania L., Melino G, & Peschiaroli A. (2022). ΔNp63-Senataxin circuit controls keratinocyte differentiation by promoting the transcriptional termination of epidermal genes. Proceedings of the National Academy of Sciences of the United States of America, 119(10), e2104718119.

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Antibody Purification and Characterization

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Antibodies used were affinity-purified rabbit IgG against the C-terminal 250 aa of Etf-1 (residues 152 to 264) (8 (link)); rabbit anti-E. chaffeensis recombinant major outer membrane proteins P28 (36 (link)); mouse monoclonal anti-HA (BioLegend); mouse monoclonal anti-GFP, anti-MnSOD, and anti-cytochrome c (Santa Cruz Biotechnology); rabbit monoclonal anti-HA (Cell Signaling Technology); rabbit anti-actin (Sigma-Aldrich); Alexa Fluor (AF)488- and AF555-conjugated goat anti-rabbit IgG and anti-mouse IgG (Life Technologies); horseradish peroxidase (HRP)-conjugated goat anti-llama IgG (Bethyl Laboratories); and HRP-conjugated goat anti-mouse IgG and HRP-conjugated goat anti-rabbit IgG (KPL).

Zhang W., Lin M., Yan Q., Budachetri K., Hou L., Sahni A., Liu H., Han N.C., Lakritz J., Pei D, & Rikihisa Y. (2021). An intracellular nanobody targeting T4SS effector inhibits Ehrlichia infection. Proceedings of the National Academy of Sciences of the United States of America, 118(18), e2024102118.

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Antibody Dilutions for Protein Detection

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1:80,000 mouse monoclonal anti-Pgk1 (22C5D8; Thermo Fisher Scientific), 1:10,000 mouse monoclonal anti-HA (901502; BioLegend), 1:10,000 mouse monoclonal anti-GFP (11814460001; Roche), 1:10,000 mouse monoclonal anti-His (51-9000012; BD), 1:10,000 rabbit polyclonal anti-MBP (E8030S; New England Biolabs, Inc.), and 1:10,000 rabbit polyclonal anti-TAP, CAB1001; Thermo Fisher Scientific) were used. 1:3,000 anti–Fab1-T1953-P is a custom rabbit polyclonal antibody raised by 21st Century Biochemicals.

Jin N., Jin Y, & Weisman L.S. (2017). Early protection to stress mediated by CDK-dependent PI3,5P2 signaling from the vacuole/lysosome. The Journal of Cell Biology, 216(7), 2075-2090.

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Yeast Protein Extraction and Western Blot

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Cells were fixed with 20% trichloroacetic acid (TCA) and disrupted by bead beating. Lysate and precipitate were mixed with 600 μL 10% TCA and pelleted. The pellet was resuspended in 1x Laemmli buffer with 5% β-mercaptoethanol and 160 mM Tris HCl (pH 7.4), boiled for 10 min and sonicated briefly. The extract was subjected to SDS gel analysis. The following antibodies were used: mouse monoclonal anti-Rad53 (clone EL7, in house [76 (link)], 1:5), mouse monoclonal anti-Pgk1 (Novex, Cat# 459250, 1:10.000), mouse monoclonal anti-HA (Biolegend, Cat# 901501, 1:10.000), rabbit polyclonal anti-Rnr3 (Agrisera. Cat# AS09574, 1:200), rabbit polyclonal anti-GST (in-house, 0.5 μg/ml), goat anti-mouse IgG (H + L)-HRP Conjugate (Bio-Rad, Cat# 1706516, 1:20000), goat anti-rabbit IgG (H + L)-HRP Conjugate (Bio-Rad, Cat# 1706515, 1:20000). Detection was performed by electrochemiluminescence (ECL, GE- Healthcare).

Ajazi A., Choudhary R., Tronci L., Bachi A, & Bruhn C. (2022). CTP sensing and Mec1ATR-Rad53CHK1/CHK2 mediate a two-layered response to inhibition of glutamine metabolism. PLoS Genetics, 18(3), e1010101.

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SDS-PAGE and Western Blot Analysis

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Proteins were solubilized in SDS sample buffer (50 mM Tris-Cl pH 6.8; 10 mM EDTA, 5% glycerol, 2% SDS, 0.01% bromophenol blue) containing 5% b-mercaptoethanol. All samples were incubated for 15 min at 65 C. For deglycosylation, SDS sample buffer solubilized proteins were diluted 1:4 in water to reduce the SDS contentration to 0.5% and were treated with EndoH (New England Biolabs) according to the manufacturer's protocol. For western blot analysis, proteins were separated by SDS-PAGE using Tris-glycine acrylamide gels and transferred to PVDF membrane followed by enhanced chemiluminescence analysis (Pierce) to detect the bound antibodies. The following primary antibodies were used: mouse monoclonal anti-FLAG (M2, Sigma; 1:1000), mouse monoclonal anti-HA (BioLegend; 1:1000) mouse monoclonal anti-b-Actin (Sigma; 1:4000). The following secondary antibodies were used: Donkey-anti-mouse IgG -HRP conjugated (Dianova; 1:10000). For detection the LAS-4000 system (Fuji) was used. Data shown are representative of three independent experiments. For quantification we used FIJI (Schindelin et al., 2012) and data acquired from the LAS-4000.

Yücel S.S., Stelzer W., Lorenzoni A., Wozny M., Langosch D, & Lemberg M.K. (2019). The Metastable XBP1u Transmembrane Domain Defines Determinants for Intramembrane Proteolysis by Signal Peptide Peptidase. Cell reports, 26(11).

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