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Goat anti rabbit igg hrp

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Goat anti-rabbit IgG-HRP is a secondary antibody that is conjugated with horseradish peroxidase (HRP) enzyme. It is used to detect and quantify rabbit primary antibodies in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry.

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Lab products found in correlation

Goat anti mouse igg hrp, by Jackson ImmunoResearch (27 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (8 mentions) Protease inhibitor cocktail, by Thermo Fisher Scientific (4 mentions) Clarity western ecl substrate, by Bio-Rad (3 mentions) Goat anti mouse igg hrp, by Santa Cruz Biotechnology (3 mentions) Sc 47778 hrp, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488 conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Hrp conjugated β actin antibody, by Santa Cruz Biotechnology (2 mentions) Rabbit monoclonal anti phospho p44 42 mapk erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions) Immun star westernc chemiluminescence kit, by Bio-Rad (2 mentions) Pvdf membrane, by Thermo Fisher Scientific (2 mentions) Rabbit anti calnexin, by Merck Group (2 mentions) Anti gfp, by Santa Cruz Biotechnology (2 mentions) Anti mouse igg hrp, by Jackson ImmunoResearch (2 mentions) Image lab 3, by Bio-Rad (2 mentions) Chemidoc xr, by Bio-Rad (2 mentions) Odyssey fc imaging system, by LI COR (2 mentions) β actin, by Proteintech (2 mentions) Anti ha, by Cell Signaling Technology (2 mentions) Anti samd14, by Cell Signaling Technology (2 mentions)

58 protocols using goat anti rabbit igg hrp

1

Western Blot Antibody Reagents

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Rabbit anti-cytosolic PEPCK was prepared in our laboratory [18 (link)]. Goat anti-muscle pyruvate kinase (PK-M) was from Rockland Immunochemicals Inc. (Gilbertsville, PA). An HRP-conjugated β-actin antibody was from Santa Cruz Biotechnology (sc-47778-HRP; Santa Cruz, CA). Rabbit monoclonal anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) was from Cell Signaling Technology Inc. (#4370, Beverly, MA). Secondary antibodies were goat anti-rabbit IgG-HRP, rabbit anti-goat IgG-HRP (Jackson ImmunoResearch, West Grove, PA), and Alexa Fluor 488-conjugated donkey anti-rabbit IgG (Molecular Probes, Eugene, OR).
Bertinat R., Westermeier F., Silva P., Gatica R., Oliveira J.M., Nualart F., Gomis R, & Yáñez A.J. (2018). The Antidiabetic Agent Sodium Tungstate Induces Abnormal Glycogen Accumulation in Renal Proximal Tubules from Diabetic IRS2-Knockout Mice. Journal of Diabetes Research, 2018, 5697970.
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2

Immunoblotting of Immunoprecipitated Proteins

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Beads containing immunoprecipitated proteins were mixed with 15 μl of 6× SDS Gel Loading Buffer, mixed and heated at 95°C for 10 minutes. The samples were then placed on the magnetic stand, allowing the beads to migrate to the magnet and the supernatant was loaded onto 15% SDS-PAG. After the electrophoresis the proteins were transferred to PVDF membrane (Thermo) for 30 min at 1 Amp and 25 V. Then the membrane was blocked in TBS containing 0.1% Tween 20 and 1% BSA at 4°C overnight. After blocking the membrane was incubated for 2 h at RT with primary IgG, washed and then incubated for 1 h at RT with HRP-conjugated secondary IgG. The following primary antibodies were used: anti-human paxillin (C-18) goat polyclonal antibody (Santa Cruz Biotechnology; 1:400), anti-human FAK (A-17) rabbit polyclonal antibody (Santa Cruz Biotechnology; 1:400), mouse anti 6xHis monoclonal antibody (Clontech; 1:500) for detection of MBP-LD His-tagged constructs after competition IP with recombinant purified FAT domain. Secondary antibodies: rabbit anti-goat IgG-HRP (Jackson ImmunoResearch; 1:2500), goat anti-rabbit IgG-HRP (Jackson ImmunoResearch; 1:2500), goat anti-mouse IgG-HRP (Jackson ImmunoResearch; 1:2500). Immun-Star WesternC Chemiluminescence Kit (BIO-RAD) was used for signal detection.
Nocula-Lugowska M., Lugowski M., Salgia R, & Kossiakoff A.A. (2015). Engineering synthetic antibody inhibitors specific for LD2 or LD4 motifs of paxillin. Journal of molecular biology, 427(15), 2532-2547.
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3

Immunoblotting with Anti-S1R, Anti-V5, and Other Antibodies

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Mouse monoclonal anti-S1R was from Santa Cruz Biotechnology. Mouse monoclonal anti-V5 was from GenScript, mouse anti-RFP, mouse anti-actin, and rabbit anti-calnexin were from Sigma. Strep-HRP, goat anti-mouse IgG-HRP, goat anti-rabbit IgG-HRP, goat anti-mouse Dylight 649, and goat anti-rabbit Dylight 488 were from Jackson-Immuno-Research Labs. Antibodies specific for peptides corresponding to the carboxyl termini of H1 were the ones used in a previous study (27 (link)).
Sharma N., Patel C., Shenkman M., Kessel A., Ben-Tal N, & Lederkremer G.Z. (2021). The Sigma-1 receptor is an ER-localized type II membrane protein. The Journal of Biological Chemistry, 297(5), 101299.
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4

Western Blot Analysis of Chondrocyte Proteins

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Total protein was isolated from the chondrocytes with different treatments using RIPA protein lysis buffer, and protein concentrations were determined using a BCA assay kit (BOSTER). The protein samples (20 μg) were separated by 10% SDS-PAGE and then transferred to PVDF membranes. After blocking with 5% skim milk at 37°C for 2 h, the membranes were incubated with anti-matrix metallopeptidase 13 (MMP13) antibody (1:4,000; Proteintech Group, Inc., Rosemont, IL, United States), anti-AKT serine/threonine kinase 1 (AKT1) antibody (1:2,000, Proteintech Group, Inc.), anti-phosphatase and tensin homolog (PTEN) antibody (1:4,000, Bioss), anti-gastrin-releasing peptide receptor (GRPR) antibody (1:2,000, Abcam), and anti-GAPDH antibody (1:2,000, Abcam) at 4°C overnight. The membranes were then incubated with the secondary antibody (goat anti-rabbit IgG-HRP, 1:1,000; Jackson ImmunoResearch Laboratories Inc., PA, United States). After incubation at 37°C for 2 h, the Millipore ECL system (Shanghai Tanon Technology Co., Ltd., Shanghai, China) was used to visualize the protein bands.
Wang X., Li Z., Cui Y., Cui X., Chen C, & Wang Z. (2021). Exosomes Isolated From Bone Marrow Mesenchymal Stem Cells Exert a Protective Effect on Osteoarthritis via lncRNA LYRM4-AS1-GRPR-miR-6515-5p. Frontiers in Cell and Developmental Biology, 9, 644380.
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5

Immune Modulation by Cytokines

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Mouse IL-4, GM-CSF, and IL-2 were purchased from Peprotech (Rocky Hill, NJ, USA). MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and CFDA-SE were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-β-actin and anti-cytochrome-C antibodies were products of Santa Cruz (Dallas, TX, USA). Rabbit-anti-human CRT polyclonal antibody was purchased from Stressgen (Victoria, BC, Canada). Goat anti-rabbit IgG-HRP was a product of Jackson (Philadelphia, PA, USA). LDM was prepared in the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences. BENS (bisethyl-norspermine) was kindly provided by Prof. Robert A. Casero at Johns Hopkins University. ELISA kits for TNF-α and IFN-γ were purchased from Boster (Wuhan, Hubei, China). Annexin V-FITC/PI apoptosis detection kit was a product of Invitrogen (San Diego, CA, USA). LDH detection kit was a product of Nanjing Jiancheng Bioengineering Institute (Nanjing, China).
Yang J.L., Qin Y., Li L., Cao C.Y., Wang Q., Li Q., Lv Y.F, & Wang Y. (2016). Apoptotic Melanoma B16-F1 Cells Induced by Lidamycin Could Initiate the Antitumor Immune Response in BABL/c Mice. Oncology Research, 23(1-2), 79-86.
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6

Comprehensive Antibody Characterization Protocol

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Mouse anti-HA tag, anti–β-COP, and anti-tubulin were from Sigma-Aldrich, and anti-GFP was from Santa Cruz Biotechnology (Dallas, TX). Rabbit anti–GM-130 was from Abcam (Cambridge, England). Mouse anti-human ERManI was a kind gift from Richard Sifers (Baylor College, Houston, TX) (used only for human cells) or was from Santa Cruz Biotechnology (used for mouse cells). Rabbit anti-calnexin was from Sigma-Aldrich, and anti-dsRED was from MBL. Goat anti-mouse immunoglobulin G (IgG) linked to horseradish peroxidase (HRP) and goat anti-rabbit IgG-HRP were from Jackson Labs (West Grove, PA). Goat anti-mouse Dylight 594 and goat anti-rabbit Dylight 488 were from Thermo Scientific (Barrington, IL). Rabbit polyclonal anti-H2 N-terminus was previously described (Tolchinsky et al., 1996 (link)), Goat anti-mouse IgG linked to agarose beads was from Sigma-Aldrich. Rabbit anti-Cab45 was described before (Scherer et al., 1996 (link)).
Benyair R., Ogen-Shtern N., Mazkereth N., Shai B., Ehrlich M, & Lederkremer G.Z. (2015). Mammalian ER mannosidase I resides in quality control vesicles, where it encounters its glycoprotein substrates. Molecular Biology of the Cell, 26(2), 172-184.
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7

Western Blot Detection of CD200 Proteins

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After transfer onto PVDF membranes, the blots were blocked with 5% milk-TBST for 1 hour at room temperature, and then probed with primary antibodies overnight at 4°C. The following primary antibodies were used in Western blotting experiments: rabbit anti-hCD200v+c serum (1:2000 dilution), rabbit anti-CD200 c-tail serum (1:500 dilution), or rabbit-hCD200R1 serum (1:2000 dilution). Regardless of the primary antibody used, following primary antibody incubation and washings in TBS-T, all blots were probed with goat anti-rabbit IgG-HRP (Jackson) at a 1:10,000 dilution for 45 minutes as secondary antibody. After thorough washing, blots were developed using an ECL Western blot detection kit (GE Healthcare Bio-Sciences).
Wong K.K., Zhu F., Khatri I., Huo Q., Spaner D.E, & Gorczynski R.M. (2016). Characterization of CD200 Ectodomain Shedding. PLoS ONE, 11(4), e0152073.
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8

Antibody Characterization for Cell Signaling Assays

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Antibodies used in this study are as follows. Anti-human Claspin was generated against the human recombinant Claspin with aa896–1,014 produced in E. coli. Anti-Chk1 phospho-S345 (#2348), anti-Chk1 phospho-S317 (#2344), anti-p44/42 MAPK (Erk1/2) (#4695), anti- SAPK/JNK (#9252), p38 MAPK (#8690), anti-p38 MAPK T180/Y182 (#4511), anti-p44/42 MAPK (Erk1/2) T202/Y204 (#4370), anti-SAPK/JNK T183/Y185 (#4668), Caspase-9 (#9508), Cleaved Caspase-3(#9661), and Mcl-1 (#5453) were obtained from Cell Signaling. Anti-α Tubulin (sc23948), anti-MCM2 (sc-9839), and anti-Chk1 (sc-8408) were obtained from Santa Cruz. Anti-phospho-H2A.X S139 (06-536) was purchased from Merck. Anti-BrdU (Ab6326) was purchased from Abcam. Anti-ATR phospho-T1989 (GTX128145) was purchased from GeneTex. Anti-BrdU (555627) was purchased from BD Pharmingen. Anti-H2A.X phospho-S139 (613402), anti-Rat IgG Alexa Fluor 555 (405420), and FITC-Anti-BrdU (364104) were purchased from Biolegend. RPA32 phospho-S4/S8 (A300-245A) and anti-MCM2 S53(A300-756A) were purchased from Bethyl. Anti-Mouse IgG Alexa Fluor 488 (A-11017) was purchased from Invitrogen. Goat Anti-Rabbit IgG HRP (111-035-003) and Goat Anti-Mouse IgG (115-035-003) were purchased from Jackson ImmunoResearch Laboratory.
Hsiao H.W., Yang C.C, & Masai H. (2023). Claspin-Dependent and -Independent Chk1 Activation by a Panel of Biological Stresses. Biomolecules, 13(1), 125.
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9

Anticancer Drug Stock Solution Preparation

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Stock solutions of NCTD (Yuanye Bio-Technology Co., Ltd., Shanghai, China), 5-FU (Selleck Biotech Co., Ltd., Houston, TX, USA), and DMSO (Sigma-Aldrich, St. Louis, MO, USA) were prepared. The preparation of NCTD and 5-FU stock solutions: 20 mg NCTD powder was diluted to 100 mM with 1.2 mL DMSO. The working concentration was 12.5/25 μM. Analogously, 20 mg 5-FU powder was diluted to 100 mM with 1.5 mL DMSO. The working concentration of 5/10 μM was then prepared. Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin/streptomycin (PS), and trypsin were obtained from Gibco, Thermo Fisher Scientific, Inc. (Waltham, MA, USA). β-actin monoclonal antibodies were purchased from Proteintech Group, Inc. (Rosemont, IL, USA). Antibodies against caspase-3 and caspase-9 were purchased from Cell Signaling Technology Inc. (Boston, MA, USA). Goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP were purchased from Jackson ImmunoResearch Inc. (West Grove, PA, USA). Cell counting kit-8 (CCK-8) was purchased from Beyotime Biotechnology (Shanghai, China). The Annexin V-FITC/PI Apoptosis Detection Kit and TUNEL FITC Apoptosis Detection Kit were purchased from Vazyme Biotech Co., Ltd. (Nanjing, China).
Huang Y., Wan X.W., Du Y.T., Feng Y., Yang L.S., Liu Y.B., Chen T., Zhu Z., Xu Y.T, & Wang C.C. (2024). Norcantharidin Enhances the Antitumor Effect of 5-Fluorouracil by Inducing Apoptosis of Cervical Cancer Cells: Network Pharmacology, Molecular Docking, and Experimental Validation. Current Issues in Molecular Biology, 46(5), 3906-3918.
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10

Immunofluorescence and Western Blot Protocols

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Anti-Marshalin21 (link) was used at 2 μg/ml for immunofluorescence (IF) and 0.6 μg/ml for Western blot. Commercial antibody dilutions were as follows: anti-V5 (Invitrogen), 1:1,000 dilution for IF and 1:5,000 for Western blot; anti-α-tubulin (Zymed, San Francisco, CA), 1:800 for IF; anti-GFP (Clontech, Mountain View, CA), 1:2,000 for Western blot; anti-GST (Sigma), 1:10,000 for Western blot; goat anti-mouse IgG-Alexa Fluor 546, goat anti-rabbit IgG-Alexa Fluor 488 or 546 (Molecular Probes, Eugene, OR), 1:500 for IF; goat anti-rabbit IgG-HRP and goat anti-mouse IgG-HRP (Jackson ImmunoResearch, West Grove, PA), 1:10,000 for Western blot; Texas Red-X phalloidin (Molecular Probes, Eugene, OR), 1:2,000 to stain F-actin. Donkey anti-mouse IgG-Alexa Fluor 647F(ab’)2 Fragment (Jackson ImmunoResearch, West Grove, PA), 1:1,000 for IF.
Takahashi S., Mui V.J., Rosenberg S.K., Homma K., Cheatham M.A, & Zheng J. (2016). Cadherin 23-C Regulates Microtubule Networks by Modifying CAMSAP3’s Function. Scientific Reports, 6, 28706.
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