Rabbit anti ha

Manufactured by Santa Cruz Biotechnology

Sourced in United States, United Kingdom

Rabbit anti-HA is a primary antibody that specifically binds to the hemagglutinin (HA) tag. The HA tag is a commonly used epitope tag that can be fused to recombinant proteins to facilitate their detection and purification. Rabbit anti-HA is a valuable tool for researchers working with HA-tagged proteins.

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Lab products found in correlation

Rabbit anti myc, by Santa Cruz Biotechnology (5 mentions) Rabbit anti flag, by Cell Signaling Technology (4 mentions) Mouse anti ha, by Santa Cruz Biotechnology (3 mentions) Mouse anti flag m2, by Merck Group (3 mentions) Mouse anti β actin, by Santa Cruz Biotechnology (3 mentions) Rabbit anti tace adam17, by Abcam (2 mentions) Mouse anti transferrin receptor, by Thermo Fisher Scientific (2 mentions) Goat anti actin, by Santa Cruz Biotechnology (2 mentions) Rabbit anti flag, by Merck Group (2 mentions) Mouse anti tubulin, by Merck Group (2 mentions) Mouse anti gfp, by Roche (2 mentions) Rabbit anti v5, by Merck Group (2 mentions) Mouse anti v5, by Thermo Fisher Scientific (2 mentions) Pierce ip lysis buffer, by Thermo Fisher Scientific (2 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Rabbit anti zo 1, by Thermo Fisher Scientific (2 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (2 mentions) Dynabeads protein g, by Thermo Fisher Scientific (2 mentions) Hrp coupled secondary antibody, by Santa Cruz Biotechnology (1 mentions) Proteinase k, by Americanbio (1 mentions)

Close spelling variants of this product

Polyclonal rabbit anti-HA (Y-11) Rabbit anti-HA (sc-805, Rabbit polyclonal anti-HA Rabbit anti-HA antibody Anti-HA rabbit polyclonal antibody Rabbit anti-HA (Y11) Rabbit HA Anti-HA Rabbit anti-TM

34 protocols using rabbit anti ha

Immunoblotting Antibodies and Detection

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The following antibodies were used: Goat anti-mouse amphiregulin (R&D Systems, catalogue number AF989; 1:1000), Mouse anti-nonphospho-Tyr1173-EGFR (Millipore, catalogue number 05-484; 1:1000), mouse anti-beta-actin (Santa Cruz, catalogue number sc-47778; 1:2000), rabbit anti-TACE/ADAM17 (Abcam, catalogue number ab39161; 1:2000), rabbit anti-HA (Santa-Cruz, catalogue number sc-805; 1:2000) and mouse anti-transferrin receptor (Invitrogen, catalogue number 13-6800; 1:1000). Corresponding species-specific HRP-coupled secondary antibodies were used from Santa Cruz and Cell Signaling.

Siggs O.M., Grieve A., Xu H., Bambrough P., Christova Y, & Freeman M. (2014). Genetic interaction implicates iRhom2 in the regulation of EGF receptor signalling in mice. Biology Open, 3(12), 1151-1157.

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Evaluating Periplasmic Protein Accessibility

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A B. thetaiotaomicron strain expressing an HA-epitope tagged allele of the periplasmic protein SusA (Shipman et al., 1999 (link)) was grown to OD600 ~0.8 in minimal media with methionine, pelleted and washed in 1x cOmplete EDTA-free protease-inhibitor cocktail (Roche, Basel, Switzerland) before being pelleted and stored at −80 ˚C. Pellets were thawed and resuspended in PBS with proteinase K (0, 10, 50 or 100 µg/mL; AmericanBio, Natick, MA, USA), and incubated at 37 ˚C aerobically under continuous agitation (250 rpm) for 8 hr. Cells were then pelleted and washed 3 times in 1x cOmplete EDTA-free protease-inhibitor cocktail, pelleted and stored at −80 ˚C. Thawed cells were lysed using BugBuster reagent (Millipore Sigma, Burlington, MA, USA), 20 µg of clarified protein lysate was loaded onto an SDS-PAGE gel, transferred to a PVDF membrane and probed with rabbit anti-BtuG2 and rabbit anti-HA (Santa Cruz Biotechnology, Dallas, TX, USA).

Wexler A.G., Schofield W.B., Degnan P.H., Folta-Stogniew E., Barry N.A, & Goodman A.L. (2018). Human gut Bacteroides capture vitamin B12 via cell surface-exposed lipoproteins. eLife, 7, e37138.

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Antibody Sources for Cell Signaling

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Mouse monoclonal anti-Flag M5 antibody was from Sigma-Aldrich. Rabbit anti-phospho-Smad2 and mouse anti-Myc were home-made and have been described before [59 (link)]. Mouse anti-HA was from Roche. Rabbit anti-HA and mouse anti-LKB1 used for co-immunoprecipitation and IHC assays, mouse anti-Smad1, rabbit anti-TGFβRI/ALK5 (V-22), rabbit anti-ID1, goat anti-STRADα, goat anti-Smad7, mouse anti-β-tubulin, mouse anti-ubiquitin (P4D1), and rabbit and mouse IgGs used for control immunoprecipitations, were from Santa Cruz Biotechnology. Mouse anti-BMPRIA/ALK3, mouse anti-BMPRIB/ALK6 and goat IgG used for control immunoprecipitations and immunoblotting, were from R&D Systems, Inc. Mouse monoclonal anti-GAPDH was from Ambion. Mouse anti-E-cadherin was from Becton Dickinson Transduction Labs. Rabbit anti-phospho-Smad1/5/8, rabbit anti-phospho-AMPK (Thr172), rabbit anti-AMPKα, rabbit anti-phospho-p70 S6 kinase (Thr389), rabbit anti-p70 S6 kinase and rabbit anti-ACVRI/ALK2 were from Cell Signaling Technology, while rabbit anti-MO25 and anti-Smad1 were from Epitomics, and were used for immunoprecipitation, immunoblot and IHC assays.

Raja E., Tzavlaki K., Vuilleumier R., Edlund K., Kahata K., Zieba A., Morén A., Watanabe Y., Voytyuk I., Botling J., Söderberg O., Micke P., Pyrowolakis G., Heldin C.H, & Moustakas A. (2015). The protein kinase LKB1 negatively regulates bone morphogenetic protein receptor signaling. Oncotarget, 7(2), 1120-1143.

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Comprehensive Viral Protein Immunodetection

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For plaques, immunostaining, and PLA, primary antibodies used were mouse anti-RSV N (Novus Biologics, Cat. No. NB100-64752 1:500 dilution), human anti-RSV F (Palivizumab, Medimmune, 1 μg/mL), mouse anti-RSV P (Abcam, Cat. No. ab94965, 1:1000 dilution), mouse anti-RSV M2-1 (Abcam, Cat, No. ab94805, 1:1000 dilution), goat anti-panRSV (Abcam, Cat. No. ab20745, 1:1000 dilution), rabbit anti-FLAG (Cell Signaling Technology, Cat. No. 14793S, 1:1000 dilution), mouse anti-V5 (Sigma Aldrich, Cat. No. V8012, 1:500 dilution), mouse anti-Myc (Cell Signaling Technology, Cat. No. 2276S, 1:1000 dilution), mouse anti-Myc-AF488 (Cell Signaling Technology, Cat. No. 2279S, 1:250 dilution), rabbit anti-HA (Santa Cruz, Cat. No. sc-805, 1:1000 dilution), and mouse anti-HA-AF488 (Novus Biologics, Cat. No. NBP2-50416AF488, 1:100 dilution). All respective secondary antibodies for immunostaining (Jackson Immunoresearch, Thermo Fisher) were used at 4 μg/ml. Secondary antibodies for PLA (Sigma Aldrich) were used according to manufacturer’s instructions.

Blanchard E.L., Braun M.R., Lifland A.W., Ludeke B., Noton S.L., Vanover D., Zurla C., Fearns R, & Santangelo P.J. (2020). Polymerase-tagged respiratory syncytial virus reveals a dynamic rearrangement of the ribonucleocapsid complex during infection. PLoS Pathogens, 16(10), e1008987.

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Western Blot Antibody Detection

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The nitrocellulose membranes were blocked and antibodies were applied in 5 % milk in TBST (100 mM TRIS–HCl, pH7.5, 0.9 % NaCl, 0.1 % Tween-20). The antibodies used were rabbit anti-G3BP1 (Millipore, 07-1801), rabbit anti-SOD1 (Santa Cruz, sc-11407), mouse anti-FLAG M2-HRP (Sigma, A8592), rabbit anti-HA (Santa Cruz, sc-805), mouse anti-PABP1 (Santa Cruz, sc-32318) and goat anti-actin (Santa Cruz, sc-1616). All immunoblotting images were acquired using a BioRad ChemiDoc MP system.

Gal J., Kuang L., Barnett K.R., Zhu B.Z., Shissler S.C., Korotkov K.V., Hayward L.J., Kasarskis E.J, & Zhu H. (2016). ALS mutant SOD1 interacts with G3BP1 and affects stress granule dynamics. Acta Neuropathologica, 132(4), 563-576.

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Immunoprecipitation and Western Blotting Analysis

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Transfected cells were lysed by rocking at 4 °C for 30 min in Pierce IP lysis buffer (Thermo Fisher Scientific) with a Roche Complete, Mini, EDTA-free protease inhibitor cocktail tablet. The lysates were subjected to immunoprecipitation with anti-Flag-conjugated agarose beads (Sigma) for 2 h at 4 °C. After washing three times with lysis buffer, immunoprecipitates were boiled in 4× NuPAGE LDS sample buffer (Novex), and western blotting was carried out according to the NuPAGE electrophoresis (Novex) protocol with rabbit anti-Flag (1:1000, Sigma), rabbit anti-MYC (1:100, Santa Cruz Biotechnology), and rabbit anti-HA (1:200, Santa Cruz Biotechnology) antibodies.
For tissue preparation, 68 hrs AEL eye discs (n = 40) were collected in cold RIPA lysis buffer (Thermo Fisher Scientific). After centrifuge at 20000 g for 10 min at 4 °C, the supernatant was transferred to a new tube and ready for western blot analysis.

Jin M, & Mardon G. (2016). Distinct Biochemical Activities of Eyes absent During Drosophila Eye Development. Scientific Reports, 6, 23228.

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Antibody Characterization for Neuronal Imaging

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Rabbit Plk2 antibodies (#7382, ICC (immunocytochemistry) 1:200, IB (immunoblotting) 1:500) used for western blotting and immunoprecipitation and guinea pig SPAR (ICC 1:400) antibodies have been described (Lee et al., 2011 (link)). The following antibodies were purchased from the indicated suppliers and used at the indicated dilutions: APP-N (Sigma A8967, ICC 1:400, IB 1:1000); RB9023 (Thermo Scientific, ICC 1:5); M3.2 (Biolegend, ICC 1:500); 4G8 (Biolegend, ICC 1:500); APP-C (Sigma A8717, ICC 1:1000); Y188 (OriGene, ICC 1:500, IB:1:1000~1:7000); C1/6.1 (Biolegend, ICC 1:400); sAPPβ (Immuno-Biological Laboratories, IB 1:500); 6E10 (Covance, ICC 1:500, IB 1:1000); 22C11 (Millipore, MAB348, IB:1:1000); mouse anti-MAP2 (Sigma, HM-2, ICC 1:400); chicken anti-MAP2 (NeuroMAb, ICC 1:400); Plk2 C-terminal C-18 and N-terminal N-17 and H-90 (Santa Cruz, ICC 1:200, IB 1:400); GFP (Invitrogen and NeuroMAb, ICC 1:200, IB 1:1000); GluA2 (BD Pharmingen, ICC 1:50~1:100); mouse anti-PSD-95 (NeuroMAb, ICC 1:200); rabbit anti-PSD-95 (Cell Signaling, ICC 1:200); guinea pig anti-Synaptotagmin 1 (Synaptic Systems, ICC 1:500); mouse anti-VGlut1 (NeuroMAb, N28/9, ICC 1:200); rabbit anti-HA (Santa Cruz, ICC 1:100); and unlabeled donkey anti-mouse IgG (Sigma). AlexaFluor-488 and AlexaFluor-555 (Invitrogen, ICC 1:200~400) were secondary antibodies used for immunocytochemistry.

Lee Y., Lee J.S., Lee K.J., Turner R.S., Hoe H.S, & Pak D.T. (2017). Polo-like kinase 2 phosphorylation of amyloid precursor protein regulates activity-dependent amyloidogenic processing. Neuropharmacology, 117, 387-400.

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Western Blot Protein Detection Protocol

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Cells were lysed in 9 M urea, 10 mM Tris-HCl pH 6.8, sonicated and clarified by centrifugation. 80 µg of protein was subjected to SDS-PAGE and transferred to nitrocellulose. Membranes were blocked in 5% non-fat dry milk in TBS-T (TBS with 0.1% Tween), then incubated with primary antibodies: mouse FLAG M2 (F1804 from Sigma; 1:5000 dilution), rabbit anti-FLAG (PA1-984B from Invitrogen; 1:5000 dilution), rabbit anti-HA (Santa Cruz 805; 1:200 dilution), rabbit anti-myc (Santa Cruz 789; 1:5000 dilution), rabbit anti-USP7 (Bethyl laboratories A300-033A; 1:10000 dilution), mouse anti-KIF20B (Santa Cruz 515194; 1:200 dilution), rabbit anti-FBXO38 (Bethyl laboratories A302-378A; 1:5000 dilution), rabbit anti-SKP1 (Santa Cruz 7163; 1:200 dilution), goat anti-actin (Santa Cruz 1615; 1:1000 dilution). Membranes were washed three times in TBS-T, followed by incubation with goat anti-mouse HRP (Santa Cruz 2005), goat anti-rabbit HRP (Sigma SAB3700878) or donkey anti-goat HRP (Sigma SAB3700285) at 1:5000 dilution. Membranes were developed using chemiluminescence reagents (ECL or ECL-prime; Santa Cruz or Amersham). Protein bands were quantified using ImageQuantTL (GE Healthcare Sciences) and normalized to actin bands.

Georges A., Coyaud E., Marcon E., Greenblatt J., Raught B, & Frappier L. (2019). USP7 Regulates Cytokinesis through FBXO38 and KIF20B. Scientific Reports, 9, 2724.

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Immunoprecipitation and Western Blotting

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Cells were lysed in NP-40 lysis buffer (50 mM Tris-HCl pH 7.4, 150 mM NaCl, 0.1% Nonidet P-40, 5 mM EDTA, 50 mM NaF, 1 mM Na₃VO₄, 10% Glycerol) supplemented with protease inhibitor mix (Sigma). For immunoprecipitation, anti-Flag M2 Affinity Gel (Sigma, A2220) was incubated with lysates containing 2 mg of proteins with rotation and washed five times with lysis buffer. Precipitated proteins were eluted by boiling the beads in Laemmli sample buffer. For Western blotting, lysates containing 30 μg of protein or immunoprecipitated proteins were separated by SDS-PAGE, transferred to nitrocellulose membranes, and probed with specific antibodies. Antibodies used for Western blotting: rabbit anti-FAM111A (Abcam, ab184572, 1:1,000); mouse anti-β-actin [clone AC-74] (Sigma, A5316, 1:5,000), rabbit anti-Flag (Cell Signaling, #2368, 1:1,000); rabbit anti-HA (Santa Cruz, sc-805, 1:1,000). Blots were imaged using ChemiDoc MP (Bio-Rad) and analyzed using the Image Lab software (version 6.0.1, Bio-Rad).

Palani S., Machida Y., Alvey J.R., Mishra V., Welter A.L., Cui G., Bragantini B., Botuyan M.V., Cong A.T., Mer G., Schellenberg M.J, & Machida Y.J. (2024). Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes. Nature Communications, 15, 2064.

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Western Blot Antibody Validation

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Commercial antibodies included rabbit anti-FLAG (Santa Cruz), rabbit anti-HA (Santa Cruz), mouse anti-tubulin (Sigma), mouse anti-HA (Cell Signaling Technology), rabbit anti-cGAS (Cell Signaling Technology), and mouse anti-STING (Proteintech). mouse anti-tubulin antibody was used for loading controls, and all mentioned antibodies were used at a dilution of 1:5,000 in 5% bovine serum albumin (BSA).

Qi S., Wang C., Zhang R., Zhu J., Hou X., Jiang Y., Li J., Ren M., Li M, & Hou F. (2022). Human STING Is Regulated by an Autoinhibitory Mechanism for Type I Interferon Production. Journal of Innate Immunity, 14(5), 518-531.

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