Goat anti rabbit igg h l

Manufactured by Cell Signaling Technology

Sourced in United States

Goat anti-rabbit IgG (H&L) is a secondary antibody that binds to rabbit primary antibodies. It is used in various immunoassays and detection methods to amplify and visualize the signal from rabbit primary antibodies.

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Lab products found in correlation

Goat anti mouse igg h l, by Cell Signaling Technology (4 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Alexa fluor 647 goat anti mouse a0473, by Beyotime (2 mentions) Tlr4 66350 1 ig, by Proteintech (2 mentions) Lipopolysaccharides l4391, by Merck Group (2 mentions) Alexa fluor 488 goat anti rabbit a0423, by Beyotime (2 mentions) Gapdh 60004 1 ig, by Proteintech (2 mentions) Anti tubulin antibody, by Merck Group (1 mentions) T9026, by Merck Group (1 mentions) Dykddddk tag d6w5b, by Cell Signaling Technology (1 mentions) Dab horseradish peroxidase color development kit, by Beyotime (1 mentions) Alcian blue solution, by Merck Group (1 mentions) 1 9 dimethylmethylene blue, by Merck Group (1 mentions) Col2a1 antibody, by Abcam (1 mentions) Phospho p65, by Cell Signaling Technology (1 mentions) Phospho iκbα, by Cell Signaling Technology (1 mentions) Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions) Recombinant human il 1β, by R&D Systems (1 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase, by Cell Signaling Technology (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions)

Close spelling variants of this product

Goat anti-rabbit IgG (H + L)-HRP conjugate HRP-conjugated goat anti rabbit IgG H + L HRP-labeled goat anti-rabbit IgG (H+L) Goat anti-rabbit IgG H&L (HRP) conjugated antibody Alexa Fluor 594-conjugated goat anti-rabbit IgG (H+L) Alexa Fluor-594 goat anti-rabbit IgG (H + L), Dylight™ 800 4XPEG-conjugated goat anti-rabbit IgG (H + L) Alexa Fluor 555-conjugated goat anti-rabbit IgG (H+L) Goat anti-Rabbit IgG (H+L)-HRP Rabbit anti-IgG

9 protocols using goat anti rabbit igg h l

Whole-Cell Lysate Preparation and Immunoblotting

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Whole-cell lysates were prepared by washing cells directly in wells with ice-cold PBS, followed by lysis on ice in RIPA buffer (50 mM Tris-HCl/pH 7.5, 1% (wt/vol) Triton X-100, 1% (wt/vol) glycerol, 0.5% sodium deoxycholate, 0.1% sodium dodecylsulphate, 137 mM sodium chloride, 1 mM sodium orthovanadate, and 0.5 mM EDTA) complemented with Complete™ protease inhibitor cocktail. Cell debris was removed by centrifugation (21,000 g, 10 min, 4°C) and the protein concentration of the supernatant was measured with BCA protein assay kit (Uptima, Montluçon, France).
LSR was detected with a rabbit polyclonal anti-LSR antibody (clone X-25) (dilution 1:500; sc-133765; Santa Cruz) and tubulin with a mouse monoclonal anti-tubulin antibody (dilution 1:10000; T9026; Sigma-Aldrich). Horseradish peroxidase-conjugated donkey anti-mouse IgG (H&L) (#610–703–124; Rockland) or goat anti-rabbit IgG (H&L) (#7074; Cell Signaling) antibodies (dilutions 1:3000) were used as secondary antibodies. Antibody signals were developed by enhanced chemiluminescence reaction.

Czulkies B.A., Mastroianni J., Lutz L., Lang S., Schwan C., Schmidt G., Lassmann S., Zeiser R., Aktories K, & Papatheodorou P. (2016). Loss of LSR affects epithelial barrier integrity and tumor xenograft growth of CaCo-2 cells. Oncotarget, 8(23), 37009-37022.

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AAV-mediated Genetic Modulation of ACE2 in HeLa Cells

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Hela cells were cultured in DMEM containing 10% FBS and 1% penicillin/streptomycin medium, and then transfected with AAV-vector, AAV-shACE2, AAV-shACE2_PD and AAV-ACE2_{(PD + Neck)} plasmids according to the manufacturer's manuals using Lipofectamine 2000 CD Transfection Reagent (Cat. No 12566014; Invitrogen, Carlsbad, CA, USA). The medium was replaced with DMEM/High Modified (-Phenol Red) after transfection for 6 h, and the cells were incubated for another 24 h for the expression of the target gene. The Amicon Ultra filtration tube (Cat. UFC801008; Merck millipore, IRL) were used to concentrate the cell supernatant for protein expression level detection. Western blot analysis was performed by standard procedure. DYKDDDDK Tag (D6W5B) Rabbit mAb antibody, GAPDH, Goat Anti-Rabbit IgG H&L and Goat Anti-Mouse IgG H&L were all purchased from Cell Signaling Technology (CST, Danvers, MA, USA).

Li M., Chen J., Liu Y., Zhao J., Li Y., Hu Y., Chen Y.Q., Sun L., Shu Y., Feng F, & Sun C. (2022). Rational design of AAVrh10-vectored ACE2 functional domain to broadly block the cell entry of SARS-CoV-2 variants. Antiviral Research, 205, 105383.

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Serum Protein Profiling in AML Patients

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Patients’ serum samples were collected before and after treatment and stored at temperatures – 70°C. The same batch of ELISA (Shanghai Jianglai Biotechnology Co., Ltd, Shanghai, China) was used to detect the levels of ICAT and β-catenin proteins in peripheral blood (PB). Before analysis, the serum samples and kit were equilibrated at room temperature (25°C) for 60 minutes (min), and standard wells and sample wells were set. After that, the assay procedure was carried out according to manufacturer’s instructions. Western blotting was used to analyze ICAT protein. The total proteins of AML samples were extracted using serum protein extraction kit (Beyotime Biotechnology Co., Ltd, Beijing, China). Protein concentration was determined by bicinchoninic acid method according to the manufacturer’s protocol (KeyGEN Biotech, China).The PVDF membranes were incubated with the ICAT antibodies (Abcam, England) at 4°C overnight, and membranes were probed with goat anti-rabbit IgG (H + L) (Cell Signaling Tech, USA), then developed by ECL substrate (Pierce, USA).

Han H., Zhu B., Xie J., Huang Y., Geng Y., Chen K, & Wang W. (2022). Expression level and prognostic potential of beta-catenin–interacting protein in acute myeloid leukemia. Medicine, 101(33), e30022.

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Inflammation and Apoptosis Analysis

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Recombinant human IL-1β was purchased from R&D (Minneapolis, MN, USA). MLT, Alcian blue solution, hydrochloride, EDTA, 1,9-dimethylmethylene blue (DMMB), and dye Hoechst 33,258 were purchased from Sigma-Aldrich (St. Louis, MO, USA). COL2A1 antibody was from Abcam (Cambridge, UK). The DAB Horseradish Peroxidase Color Development Kit was from (Beyotime Biotechnology, Beijing, China), and the MEBSTAIN Apoptosis TUNEL Kit Direct was from MBL International Co. (Woburn, MA, USA). The subsequent antibodies (Abs) were bought from Cell Signaling Technology (CST, Danvers, MA, USA): P65, phospho-P65, IκBα, phospho-IκBα, GAPDH, goat anti-rabbit IgG H&L (HRP), and goat anti-mouse IgG H&L (HRP).

Gao B., Gao W., Wu Z., Zhou T., Qiu X., Wang X., Lian C., Peng Y., Liang A., Qiu J., Zhu Y., Xu C., Li Y., Su P, & Huang D. (2018). Melatonin rescued interleukin 1β-impaired chondrogenesis of human mesenchymal stem cells. Stem Cell Research & Therapy, 9, 162.

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CAR T Cell Activation Signaling

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A total of 1×10⁷ CAR T cells per CAR were stimulated with 3T3-hCD19 target cells for 24 hours or left unstimulated. Cells were lysed with radioimmunoprecipitation assay buffer and total protein was quantified by ND1000 (Nanodrop); 20 µg of lysate was loaded onto the gel. Proteins were detected using anti-phospho-lymphocyte-specific protein tyrosine kinase (pLCK) (1:1000 5% BSA; phospho Tyr505; Cell SignaliNg), anti-lymphocyte-specific protein tyrosine kinase (LCK) (1:2000 5% BSA; BD), and anti-glyceraldehyde 3-phosphate dehydrogenase (1:5000 5% BSA; Cell SignaLing). Goat antirabbit IgG (H+L) and goat antimouse IgG (H+L) (Cell Signaling) were used as secondary antibodies. The secondary antibodies were diluted in 5% BSA at a 1:10 000 dilution. Primary antibodies were incubated at 4°C overnight, and secondary antibodies were incubated at room temperature for 2 hours. ChemiDocMP Imaging System (Bio-Rad) was used to detect enhanced chemiluminescence (Pierce ECL Western Blotting Substrate) western blotting signals.

Roselli E., Boucher J.C., Li G., Kotani H., Spitler K., Reid K., Cervantes E.V., Bulliard Y., Tu N., Lee S.B., Yu B., Locke F.L, & Davila M.L. (2021). 4-1BB and optimized CD28 co-stimulation enhances function of human mono-specific and bi-specific third-generation CAR T cells. Journal for Immunotherapy of Cancer, 9(10), e003354.

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Immunofluorescence Assay for Protein Expression

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Cells were fixed in freshly prepared 4% paraformaldehyde for 20 minutes at room temperature, washed with PBS and subsequently blocked and permeabilized with 0.1% Triton X in 10% goat serum containing PBS in room temperature for 1 hour. Cells were then probed with primary antibodies, Ki-67 1:500 dilution (D2H0 rabbit mAb, Cell Signaling), E6 1:100 dilution (mouse anti-virus, clone C1P5, Invitrogen), E7 1:100 dilution (mouse anti-virus, clone TVG701Y, Invitrogen) diluted in 10% goat-serum PBS and incubated overnight at 4 degree Celsius. Cells were washed with PBS and then probed with secondary antibody, goat anti-rabbit IgG (H+L), Fab2 Alexa Fluor 594, goat anti-mouse IgG (H+L), or Fab2 Alexa Fluor 488 (Cell Signaling) at 1:400 dilution in 2% goat serum containing PBS for 1 hour at room temperature followed by PBS washes and mounted with DAPI (Fluoroshield, Sigma). Imaging was completed using Fluorescence Eclipse Ti2 Inverted microscope (Nikon).

Miller D.N., Bryant J.E., Madsen E.L, & Ghiorse W.C. (1999). Evaluation and Optimization of DNA Extraction and Purification Procedures for Soil and Sediment Samples. Applied and Environmental Microbiology, 65(11), 4715-4724.

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Antibody-Based Signaling Pathway Analysis

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Antibodies: AKT (4691), P38 (8690), ERK1/2 (4695), p-AKT (4060), p-P38 (4511), p-ERK1/2 (4370), IκBα (4814), p-IκBα (2859), p-IKKα/β (2697), NF-κB (8242), Rab5a (46449), horse anti-mouse IgG (H&L) (7076), and goat anti-rabbit IgG (H&L) (7074) were purchased from Cell Signaling Technology (Danvers, MA, United States). IKKβ (ab124957), PBR (ab109497) was from Abcam (CA, United States). Alexa Fluor 488 goat anti-rabbit (A0423) and Alexa Fluor 647 goat anti-mouse (A0473) secondary antibodies were obtained from Beyotime Biotechnology (Shanghai, China). TLR4 (66350-1-Ig), GAPDH (60004-1-Ig), and β-Tubulin (10068-1-AP) were purchased from Protein-Tech (Wuhan, Hubei, China). Lipopolysaccharides(L4391) was obtained from Sigma (Louis, MO, United States), Remimazolam Tosilate for Injection (Remimazolam) were obtained from Hengrui (Jiangsu, China).

Liu X., Lin S., Zhong Y., Shen J., Zhang X., Luo S., Huang L., Zhang L., Zhou S, & Tang J. (2021). Remimazolam Protects Against LPS-Induced Endotoxicity Improving Survival of Endotoxemia Mice. Frontiers in Pharmacology, 12, 739603.

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Western Blot Analysis of Cell Signaling

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Western blot was performed as described elsewhere [7 (link)]. Total cell lysates from cultures, cocultures, or immunoprecipitated samples were resolved by 7.5% SDS–PAGE. Molecular weight markers were precision plus protein dual-color standards, BIO-RAD catalog #161-0374. Proteins were transferred to nitrocellulose membranes. Membranes were blocked with 5% nonfat dry milk in TBS-T for 1 h at room temperature and then incubated overnight at 4 °C with the corresponding antibodies: rabbit anti-Gal lectin (0.4 μg/mL made in our laboratory) [6 (link)], goat anti-human c-Met receptor (0.2 μg/mL SIGMA H9786), or goat anti-human HGF (0.2 μg/mL SIGMA H7157). Membranes were washed with TBS-T and incubated with goat anti-rabbit IgG (H + L) (1/60,000 Cell signaling 7074) or bovine anti-goat IgG (H + L) (1:10,000 Santa Cruz Biotechnology sc-2350), all conjugated to horseradish peroxidase for 90 min at room temperature. After washing with TBS-T, antibody-reactive proteins were detected by chemiluminescence.

Pérez-Hernández J., Retana-González C., Ramos-Martínez E., Cruz-Colín J., Saralegui-Amaro A., Baltazar-Rosario G., Gutiérrez-Ruíz C., Aristi-Urista G, & López-Vancell R. (2021). Entamoeba histolytica Trophozoites Interact with the c-Met Receptor at the Surface of Liver Origin Cells through the Gal/GalNAc Amoebic Lectin. Life, 11(9), 923.

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Investigating Cell Signaling Pathways

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Antibodies AKT (4691), p-AKT (4060), P38 (8690), p-P38 (4511), ERK1/2 (4695), p-ERK1/2 (4370), IkBa (4814), p-IkBa (2859), p-IKKa/b (2697), NF-kB (8242), Rab5a (46449), horse anti-mouse IgG (H&L) (7076) and goat anti-rabbit IgG (H&L) (7074) were obtained from Cell Signaling Technology (Danvers, MA, USA). IKKβ (ab124957), PBR (ab109497) was from Abcam (CA, USA). Alexa Fluor 488 goat anti-rabbit (A0423) and Alexa Fluor 647 goat anti-mouse (A0473) secondary antibodies were obtained from Beyotime Biotechnology (Shanghai, China). TLR4 (66350-1-Ig), GAPDH (60004-1-Ig) and b-Tubulin (10068-1-AP) were obtained from Proteintech (Wuhan, Hubei, China). Lipopolysaccharides(L4391) was from Sigma (Louis, MO, USA), Remimazolam Tosilate for Injection (Remimazolam)were obtained from Hengrui (Jiangsu, China).

Liu X., Lin S., Zhong Y., Shen J., Zhang X., Luo S., Zhang L., Zhou S., & Tang J. (2021). Remimazolam protects against LPS-induced endotoxicity improving survival of endotoxemia mice.

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