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Anti rabbit igg hrp

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Anti-rabbit IgG-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and bind to rabbit primary antibodies in various immunoassays and immunodetection techniques.

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Lab products found in correlation

Anti mouse igg hrp, by Jackson ImmunoResearch (20 mentions) Clarity western ecl substrate, by Bio-Rad (2 mentions) Chemidoc, by Bio-Rad (2 mentions) Mouse anti flag, by Merck Group (2 mentions) Flag tag (flag), by Merck Group (2 mentions) Mouse monoclonal anti flag m2 antibody, by Merck Group (2 mentions) Mouse monoclonal anti v5 antibody, by Thermo Fisher Scientific (2 mentions) Nitrocellulose membrane, by Carl Roth (1 mentions) Immobilon, by Merck Group (1 mentions) Protease inhibitor mix, by Thermo Fisher Scientific (1 mentions) Anti human igg hrp, by Southern Biotech (1 mentions) Rat anti ha 3f10, by Roche (1 mentions) Anti rat igg hrp, by Jackson ImmunoResearch (1 mentions) Anti mouse igg hrp, by Cell Signaling Technology (1 mentions) Nupage 4 12 bis tris protein gel, by Thermo Fisher Scientific (1 mentions) Chemidoc touch imaging system, by Bio-Rad (1 mentions) Lds gel loading buffer, by Thermo Fisher Scientific (1 mentions) Mouse anti flag m2, by Merck Group (1 mentions) Turboblot, by Bio-Rad (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Rabbit anti-HRP (12 citations) Rabbit anti-IgG (2 citations) Rabbit IgGs (2 citations)

33 protocols using anti rabbit igg hrp

1

Western Blot Analysis of FcRn and β-Actin

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The Western blot was performed as described before [36 (link)]. Briefly, the cell and tissue lysates with equal weight or cell count were extracted by using ice-cold RIPA cell lysis (10 mM Tris-Cl, pH 8.0; 1 mM EDTA, 0.5 mM EGTA, 1% Triton X-100; 0.1% sodium deoxycholate, 0.1% SDS, 140 mM NaCl; protease inhibitor mix (Thermo Fisher Scientific, Dreieich, Germany) and ultrasound (Hielscher, Stuttgart, Germany)). Equal volumes of homogenized tissue were loaded, separated in a 12.5% SDS PAGE and blotted onto a nitrocellulose membrane (Carl Roth, Karlsruhe, Germany). The membrane was blocked with 5% skimmed milk powder (TSI GmbH, Zeven, Germany) in PBS-0.1% Tween20, pH 7.4 for 1 h at room temperature (RT). Primary antibodies against FcRn [14 (link)]; for human: Sigma Aldrich, Taufkirchen, Germany) and against β-Actin (Sigma Aldrich, Taufkirchen, Germany) were diluted 1:5000 in blocking buffer and incubated overnight at 4 °C. Secondary antibodies were diluted 1:5000 (Anti-human IgG-HRP, SouthernBiotech, Birmingham, AL, USA), 1: 50000 (Anti-rabbit IgG-HRP, Jackson Immuno Research, Ely, UK) and 1:4000 (Anti-murine IgG–HRP, Sigma Aldrich, Taufkirchen, Germany). For signal detection, the chemiluminescence substrate Immobilon® (Merck Millipore, Darmstadt, Germany) was used according to the manufacturer’s instructions.
Ladel S., Maigler F., Flamm J., Schlossbauer P., Handl A., Hermann R., Herzog H., Hummel T., Mizaikoff B, & Schindowski K. (2020). Impact of Glycosylation and Species Origin on the Uptake and Permeation of IgGs through the Nasal Airway Mucosa. Pharmaceutics, 12(11), 1014.
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2

Western Blot Analysis of Protein Targets

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Proteins extracts were separated by SDS-PAGE and transferred onto a nitrocellulose membrane. The following primary antibodies were used: rabbit anti-Top2 (T. Hsieh and D. Ardnt-Jovin) (1:2000), rat anti-Vasa (DSHB) (1:200), mouse anti-actin (DSHB) (1:100), rat anti-HA 3F10 (Roche) (1:2000). The secondary antibodies were anti-mouse IgG HRP (Jackson Laboratories) (1:2000), anti-rabbit IgG HRP (Jackson Laboratories) (1:2000), and anti-rat IgG HRP (Jackson Laboratories) (1:2000).
Bhargava V., Goldstein C.D., Russell L., Xu L., Ahmed M., Li W., Casey A., Servage K., Kollipara R., Picciarelli Z., Kittler R., Yatsenko A., Carmell M., Orth K., Amatruda J.F., Yanowitz J.L, & Buszczak M. (2019). GCNA preserves genome integrity and fertility across species. Developmental cell, 52(1), 38-52.e10.
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3

Co-immunoprecipitation and Western Blotting

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Co-immunoprecipitated fractions taken from solubilized cell lysate (L), unbound waste (U), bead wash (W), and final elution (E) were mixed with LDS gel loading buffer (Thermo-Fisher, #NP0007) containing 50mM fresh dithiothreitol and heated to 65°C for 5 minutes. Samples were then run on NuPage 4–12% Bis-Tris Protein Gels (Thermo-Fisher, #NP0322) according to manufacturer’s instructions. Samples were then transferred to Immobilon PVDF membranes for Western blotting (Sigma-Aldrich, #IPSN07852) according to manufacturer’s instructions. Protein-adhered membranes were blocked with TBS-T (50mM Tris, 150mM NaCl, 0.1% Tween 20) with 3% BSA for 1 hr followed by overnight incubation with primary antibodies to detect mVenus (Anti- GFP, rabbit, 1:1000 dilution, Novus Biologicals, #NB600–308) and FLAG (Anti-FLAG- M2, mouse, 1:1000, Sigma-Aldrich, F1804) at 4°C. The following day membranes were washed 4 × 15 min in TBST and probed for 1 hr with anti-rabbit IgG HRP (1:5000, Jackson ImmunoResearch, #711-035-152) for mVenus detection or anti-mouse IgG HRP (1:3000, Cell Signaling, #7076S) for FLAG detection. Blots were washed again 4× 15 min in TBST, mixed with Clarity Western ECL Substrate (BioRad, #1705061) and imaged on a ChemiDoc Touch Imaging System (BioRad, #1708370).
English J.G., Olsen R.H., Lansu K., Patel M., White K., Cockrell A.S., Singh D., Strachan R.T., Wacker D, & Roth B.L. (2019). VEGAS as a Platform for Facile Directed Evolution in Mammalian Cells. Cell, 178(3), 748-761.e17.
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4

Western Blotting of Mouse Sperm Proteins

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Whole sperm protein was extracted as previously described (66 (link), 67 (link), 68 ). In short, mouse epididymal spermatozoa (1 × 106 cells) washed in PBS were directly lysed in 20 μl 2× SDS sample buffer or 8 M urea. The whole sperm lysate was centrifuged at 15,000g, 4 °C for 10 min. Supernatant were adjusted to 50 mM DTT and denatured at 95 °C for 10 min before loading to a SDS-PAGE gel. The proteins were transferred on a nitrocellulose membrane. The membrane was incubated overnight at 4 °C with primary antibody, followed by washing with PBST and incubation with appropriate secondary antibody for 1 h. Bands were visualized using chemiluminescence and imaged by ChemiDoc (Bio-Rad). Primary antibodies used for Western blotting were rabbit anti-mouse TXNRD3 (1:2000), GPX4 (1:2000), ANT4 (1 μg/ml), P2X2 (1 μg/ml), TOM20 (0.2 μg/ml), TXNRD1 (1 μg/ml), TXNRD2 (1 μg/ml), and mouse anti-acetylated tubulin (1: 20,000). Secondary antibodies were anti-rabbit IgG-HRP (1:10,000) and anti-mouse IgG-HRP (1:10,000) from Jackson ImmunoResearch. Normalized expression was measured by ImageJ software (v1.53).
Wang H., Dou Q., Jeong K.J., Choi J., Gladyshev V.N, & Chung J.J. (2022). Redox regulation by TXNRD3 during epididymal maturation underlies capacitation-associated mitochondrial activity and sperm motility in mice. The Journal of Biological Chemistry, 298(7), 102077.
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5

Quantification and Immunoblotting Techniques

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Total protein lysate was quantified using a standard Bradford assay and 10 μg of lysate was used for immunoblotting experiments. For Blue Native PAGE, cells were lysed in 1x Sample Preparation buffer (ThermoFisher) containing 1% digitonin. 1% SDS was supplemented for SDS-PAGE. All proteins were transferred to PVDF membranes using TurboBlot (Bio-rad) at 2.5 mA for seven mins. The primary antibodies used were anti-FLAG (M2, Sigma-Aldrich Cat# F3165, RRID:AB_259529), anti-GAPDH (Santa Cruz Biotechnology Cat# sc-47724, RRID:AB_627678) and anti-HA (Y-11, Santa Cruz Biotechnology Cat# sc-805, RRID:AB_631618). The secondary antibodies were anti-rabbit IgG-HRP (Jackson ImmunoResearch Labs Cat# 111-035-003, RRID:AB_2313567), anti-mouse IgG-HRP (light chain specific) (Jackson ImmunoResearch Labs Cat# 205-032-176, RRID:AB_2339056) and anti-mouse IgG-HRP (Jackson ImmunoResearch Labs Cat# 115-035-003, RRID:AB_10015289).
Chia P.H., Zhong F.L., Niwa S., Bonnard C., Utami K.H., Zeng R., Lee H., Eskin A., Nelson S.F., Xie W.H., Al-Tawalbeh S., El-Khateeb M., Shboul M., Pouladi M.A., Al-Raqad M, & Reversade B. (2018). A homozygous loss-of-function CAMK2A mutation causes growth delay, frequent seizures and severe intellectual disability. eLife, 7, e32451.
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6

Immunoblotting of Shigella Isogenic Strains

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M90T WT, ipaB4, ipaB4 ΔipgC, and ipaB4 ΔmxiE isogenic strains harboring either one of plasmids pNS1-8 (Table S3) were inoculated in TSB supplemented with ampicillin (100 μg/mL) and incubated overnight at 37°C with shaking at 250 rpm. Next, the resulting cultures were diluted 1:1 in laemmli 2× and heated at 95°C for 5 min and ran into 4 to 15% polyacrylamide gradient SDS-PAGE gels (Bio-Rad, number 456-8086). Immunoblotting was performed as described in (13 (link)), using the following antibodies as indicated: primary antibodies: 1/5000 mouse anti-FLAG (Sigma. number F3165), 1/10000 rabbit anti-IpaH, and 1/1000 mouse anti-RecA (MBL, number ARM191); secondary antibodies: 1/25000 (except for RecA 1/10000) anti-Mouse IgG-HRP (Jackson Immunoresearch, number 115-035-003), and 1/10000 anti-Rabbit IgG-HRP (Jackson Immunoresearch, number 111-035-003).
Silué N, & Campbell-Valois F.X. (2022). icaR and icaT are Ancient Chromosome Genes Encoding Substrates of the Type III Secretion Apparatus in Shigella flexneri. mSphere, 7(3), e00115-22.
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7

Comparative Analysis of HIF-1α and HIF-2α

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The expression of HIF-1α and HIF-2α was compared in non-pigmented and pigmented melanoma cells, with Caki cells as a positive control for HIF-2α [60 (link)], and Hif1a wild type (WT) and knockout (KO) mammary tumor epithelial cells (MTECs) derived from the polyoma virus middle T transgenic mouse (MMTV-PyMT) [61 (link)] as controls for HIF-1α. Cells were grown to 80% confluence and harvested after culture for 6 hours at either normoxia (ambient air; 5% CO2) or hypoxia (0.5% O2, 5% CO2). Cell extracts were prepared using a modified RIPA buffer to produce a whole cell extract (WCE) comprised of cytoplasmic and nuclear proteins. Insoluble material remaining after preparation of WCE was then re-extracted in high-salt (HS) (400 mM NaCl) buffer containing the deubiquitinase inhibitor N-ethylmaleimide (NEM, 0.5 μM) to enrich for nuclear proteins. HS-WCE was resolved on 3% to 8% Tris-acetate gels (10 μg/lane; Invitrogen) and transferred onto polyvinylidene fluoride (PVDF) membrane. Membranes were blocked in 5% milk prior to addition of anti-HIF1A (1:5,000, Novus Biologicals, NB100-479) or anti-HIF2A (1:2,000, Novus Biologicals, NB100-122) primary antibodies, which were incubated overnight at 4°C followed by incubation in anti-rabbit igG-HRP (Jackson Laboratories, 1;40,000) for 45 minutes and detection using enhanced chemiluminescence (ECL).
Slominski A., Kim T.K., Brozyna A.A., Janjetovic Z., Brooks D., Schwab L., Skobowiat C., Jozwicki W, & Seagroves T. (2014). The role of melanogenesis in regulation of melanoma behavior: Melanogenesis leads to stimulation of HIF-1α expression and HIF-dependent attendant pathways. Archives of biochemistry and biophysics, 563, 79-93.
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8

Immunoblotting Technique for Protein Analysis

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For immunoblotting, cells were harvested and lysed in Triton X-100 lysis buffer [20 mM Tris-HCl pH 8.0, 1% Triton X-100, 10% Glycerol, 150 mM NaCl, protease inhibitor cocktail (Roche)]. Lysates were then loaded on a 10% SDS-PAGE followed by transfer on a polyvinylidene difluoride membranes (BioRad, Mississauga, ON, Canada). Membranes were blocked in 5% dry milk TBS-T (TBS, 0.1% Tween-20) for 2 h at room temperature. Primary antibodies against mouse IRAK1 and β-actin (Cell Signaling Technologies) were diluted in TBS-T and incubated with the membranes o/n at 4°C. Antirabbit IgG HRP (Jackson ImmunoResearch, West Grove, PA, USA) were used as secondary antibodies whereby they were diluted in TBS-T and incubated with the membranes for 1 h at room temperature. Detection was performed with ECL chemiluminescence kit (BioRad).
Chartrand K., Lebel M.È., Tarrab E., Savard P., Leclerc D, & Lamarre A. (2018). Efficacy of a Virus-Like Nanoparticle As Treatment for a Chronic Viral Infection Is Hindered by IRAK1 Regulation and Antibody Interference. Frontiers in Immunology, 8, 1885.
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9

EGFR Signaling Pathway Analysis

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Antibodies and reagents were purchased as follows: mAb-2E9 (Abcam ab8465, RRID: AB_2096462), Anti-EGFR Affibody® Molecule (Abcam ab95116; RRID: AB_11156238), Anti-EGFR (D38B1, Cell Signaling Technology (CST) 4267; RRID: AB_2246311), Anti-Phospho-Akt (Ser473, D9E, CST 4060; RRID: AB_2315049), Anti-beta-Actin Monoclonal Antibody, HRP Conjugated (13E5 CST 5125; RRID: AB_1903890), Anti-Phospho-EGF Receptor (Tyr1068, D7A5 CST 3777; RRID: AB_2096270), Anti-EGFR (R & D Systems, AF231; RRID: AB_355220), Anti-EGFR (phospho Y992, EM-12, Abcam ab81440; RRID: AB_1658463), Anti-mouse IgG-HRP (Jackson ImmunoResearch 715-035-150; RRID: AB_2340770), Anti-rabbit IgG-HRP (Jackson ImmunoResearch 711-035-152; RRID: AB_10015282), Anti goat IgG-HRP (Jackson ImmunoResearch 705-035-147; RRID: AB_2313587), Anti-Rabbit-HRP (Dako P044801, RRID: AB_2617138), Recombinant Murine EGF (Peprotech, 315-09), Recombinant Murine IL-3 (Peprotech, 213-13), Erlotinib Hydrochloride (Biovision, 1558-100), Lapatinib Ditosylate (Biovision, 1642-25), 4% Paraformaldehyde (EM Grade, Electron Microscopy Service (EMS), 157-4), 25% Glutaraldehyde (Grade I, Sigma G5882), FluoSpheres™ (carboxylate modified, 0.1 μm, infrared (715/755), Invitrogen F8799).
Iyer R.S., Needham S.R., Galdadas I., Davis B.M., Roberts S.K., Man R.C., Zanetti-Domingues L.C., Clarke D.T., Fruhwirth G.O., Parker P.J., Rolfe D.J., Gervasio F.L, & Martin-Fernandez M.L. (2024). Drug-resistant EGFR mutations promote lung cancer by stabilizing interfaces in ligand-free kinase-active EGFR oligomers. Nature Communications, 15, 2130.
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10

Western Blot Analysis of Apoptosis Markers

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T cells with a >95% purity were purified using an EasySep mouse T cell enrichment kit (StemCell Technologies). T cell lysates were prepared in sample buffer (50 mM Tris-Cl [pH 6.8], 50mM 2-ME, 2% SDS, 0.2% bromophenol blue, and 10% glycerol). Antibodies used for Western blots were rabbit anti-LC3 (polyclonoal P014, MBL International), hamster anti-Bcl-2 (polyclonal, BD pharmingen), rabbit anti-Bcl-xL (polyclonal), rabbit anti-Mcl-1 (polyclonal, Rockland Immunochemicals), rabbit anti-Bim (polyclonal, Cell Signaling), rabbit anti-Bax (polyclonal, Cell Signaling), rabbit anti-Bak (polyclonal, Cell Signaling), rabbit anti-Bid (polyclonal, Abcam), rabbit anti-PARP-1 (polyclonal, Cell Signaling), rabbit anti-COX IV (polyclonal, Cell Signaling), mouse anti-cytochrome c (7H8.2C12, BioLegend), mouse anti-α-Tubulin (B-5-1-2, Sigma) and goat anti-β-Actin (polyclonal, Santa Cruz Biotechnology). For HRP-labeled western blot, the secondary antibodies were anti-rabbit IgG-HRP, anti-mouse IgG-HRP, anti-hamster IgG-HRP and anti-goat IgG-HRP (Jackson Immunoresearch). The development of the western blot was achieved with SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific). For fluorescent western blot, the secondary antibodies were anti-rabbit IgG-Alexa Fluor 680, anti-mouse IgG-Alexa Fluor 680, and anti-goat IgG-Alexa Fluor 790 (Molecular Probes, Invitrogen).
He M.X, & He Y.W. (2015). c- FLIP protects T lymphocyte from apoptosis in the intrinsic pathway. Journal of immunology (Baltimore, Md. : 1950), 194(7), 3444-3451.
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