Goat anti rabbit igg h l hrp

Manufactured by Affinity Biosciences

Sourced in United States, China

Goat anti-rabbit IgG (H+L) HRP is a secondary antibody that binds to rabbit primary antibodies. It is conjugated with horseradish peroxidase (HRP), which can be used for detection in various immunoassay applications.

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Lab products found in correlation

26 protocols using goat anti rabbit igg h l hrp

Western Blot Analysis of Tight Junction Proteins

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The tissue samples were homogenized with RIPA buffer containing phosphatase inhibitors and protease inhibitors (#sc-364162, Santa Cruz, CA, USA) to extract the total protein. All protein samples were assigned to two pieces of 6% or 10% SDS-PAGE gel (#P0012AC, Beyotime Biotechnology, Jiangsu, China) separately and then transferred to PVDF membranes (#IPVH00010, Millipore, MA, USA). The processes were performed in same tank simultaneously. After blocking and subsequent washing, primary and second antibody incubation was carried out successively, and the target proteins on the membrane were observed by the FluorChem FC2 system (Alpha, Germany).
The primary antibodies used were as follows: ZO-1 (#AF5145, Affinity, OH, USA), occludin (#DF7504, Affinity), AMPK (#AF6423, Affinity), p-AMPK (#AF3423, Affinity), CDX2 (#DF7606, Affinity), β-actin (#AF7018, Affinity). The secondary antibodies, goat anti-rabbit IgG (H+L) HRP (#S0001, Affinity,), were also used.

Wang J., Zhang Q., Xia J, & Sun H. (2022). Moderate Treadmill Exercise Modulates Gut Microbiota and Improves Intestinal Barrier in High-Fat-Diet-Induced Obese Mice via the AMPK/CDX2 Signaling Pathway. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy, 15, 209-223.

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Quantifying Apoptotic Proteins in Retinal Cells

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According to our previous studies, total protein was extracted from retinal tissues or RGCs using lysis buffer (Zhang et al. 2017a (link), b (link)). Protein concentrations were measured using a NanoPhotometer N50 Touch spectrophotometer (IMPLEN, Heidelberg, Germany). Equal amounts of protein per lane (30 µg) were run on a 10% SDS–PAGE gel and then transferred to polyvinylidene difluoride membranes. The blots were probed with primary antibodies against caspase-3 ([1:1000], 9662; CST, Danvers, MA, US), cleaved caspase-3 ([1:1000], 9661; CST, Danvers, MA, US), Tle4 (15140, [1:1500]; Novusbio, Minneapolis, MN, US), and β-actin (66009-I-Ig, [1:1000]; Proteintech, Rosemont, IL, US) followed by incubation with secondary antibodies (goat anti-rabbit IgG (H + L) HRP [1:5000] or goat anti-mouse IgG (H + L) HRP [1:5000], Affinity, Cincinnati, OH, US). The expression of target proteins was normalized to the corresponding β-actin expression level in the same sample and quantified using ImageJ software (National Institutes of Health, Bethesda, MD, USA). Each immunoblot was repeated three times to confirm the results.

Feng Y., Lu J., Peng X., Ge Y., Zhang R, & Li H. (2023). Long noncoding RNA uc007nnj.1 mediates neuronal death induced by retinal ischemia/reperfusion in mice via the miR-155-5p/Tle4 axis. Molecular Medicine, 29, 9.

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Curcumin and Chondroitin Sulfate Ameliorate MIA-Induced Osteoarthritis

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A food supplement of curcumin was provided by Chr. Hansen Holding A/S (Batch: PO100790, Hoers Holm, Denmark). Food supplement of chondroitin sulfate (Chondroitin sulfate Na, 38.80%, derived from a novel enzymatic hydrolyzed chicken sternal cartilage extract) was obtained from Meitek Technology Co., Ltd (CJ19101003, Qingdao, China). MIA was purchased from Sigma-Aldrich (St. Louis, MO, United States); EDTA decalcification solution (quick decalcification) was purchased from Servicebio (Wuhan, China); enzyme-linked immunosorbent assay (ELISA) kits of TNF-α, IL-1β, SOD, CAT, MMP-3, COL-II, PGE2, and COMP were obtained from Jiangsu Meimian Industrial Co., Ltd. (Nanjing, China). NF-kappaB and p-NF-kappaB antibodies were purchased from Cell Signaling Technology (MA, United States). cox-2, TLR4, and GAPDH antibodies were purchased from Signalway Antibody Co., Ltd. (MD, United States). Goat anti-mouse IgG (H + L) HRP and goat anti-rabbit IgG (H + L) HRP were purchased from Affinity Biosciences (OH, United States).

Guan T., Ding L.G., Lu B.Y., Guo J.Y., Wu M.Y., Tan Z.Q, & Hou S.Z. (2022). Combined Administration of Curcumin and Chondroitin Sulfate Alleviates Cartilage Injury and Inflammation via NF-κB Pathway in Knee Osteoarthritis Rats. Frontiers in Pharmacology, 13, 882304.

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Immunohistochemical Analysis of TLR2 in Liver

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The tissues were sliced with the thickness of 5 μm sections with routine deparaffinization. The sections were incubated with rabbit-anti-TLR2 antibody, rabbit-anti-α-SMA, rabbit-anti-CK-19 (Abcam, Cambridge, USA), Goat-anti-rabbit IgG H&L (HRP) (Affinity bioscience, Jiangsu, China) and DAB staining (Solarbio, Beijing, China). Details of the antibodies were presented in Table 1. The expression levels of TLR2 in liver were digitized and analyzed using Image-Pro Plus software.

Wang Y., Zhang X., Wang X., Zhang N., Yu Y., Gong P., Zhang X., Ma Y., Li X, & Li J. (2023). Clonorchis sinensis aggravates biliary fibrosis through promoting IL-6 production via toll-like receptor 2-mediated AKT and p38 signal pathways. PLOS Neglected Tropical Diseases, 17(1), e0011062.

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Neuroinflammation and Synaptic Dysfunction Markers

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KCC2 (Millipore, 07-432), GABA A R (Abcam, ab94585), GABA B R (Affinity, AF0162), BDNF (Affinity, DF6387), β-Amyloid 1-42 (Abcam, ab201060), TNF-α(Affinity, AF7014), IL-6 (Servicebio, GB11117), IL-1β (Affinity, AF5103), NLRP1 (Abcam, ab3683), NLRP3 (Abcam, ab214185), AMPAR (Abcam, ab31232), PSD95 (Thermo, MA1-045), Iba-1 (Servicebio, GB11105), GFAP (Servicebio, GB12096), β-Tubulin (Affinity, T0023), β-Actin (Affinity, AF7018), Goat Anti-Rabbit IgG (H+L) HRP (Affinity, S0001), Goat Goat Anti-Mouse IgG (H+L) HRP (Affinity, S0002).

Zhou Y., Cheng Y., Li Y., Ma J., Wu Z., Chen Y., Mei J, & Chen M. (2021). Soluble β-amyloid impaired the GABA inhibition by mediating KCC2 in early APP/PS1 mice. Bioscience trends, 15(5).

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Protein Extraction and Western Blot Analysis

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Tissues were homogenized in RIPA buffer supplemented with protease and phosphatase inhibitors (NCM Biotech, China), sonicated to shear DNA, and centrifuged at 14,000× g for 20 min at 4°C. Equal amounts of protein sample (50 μg) were resolved by SDS-PAGE. The following primary antibodies were used: rabbit anti-β-actin antibody (1:10,000; Affinity, Cincinnati, OH), rabbit anti-bace1 antibody (1:1,000; Cell Signaling Technology, Boston, MA), rabbit anti-NF-KB antibody (1:1,000; Cell Signaling Technology, Boston, MA), and goat anti-rabbit IgG (H+L) HRP (1:5,000; Affinity, Cincinnati, OH). Quantification of immunoreactive bands is reported as a ratio against β-actin using ImageJ software.

Luo S.M., Li L.Y., Guo L.Z., Wang L., Wang Y.F., Chen N, & Wang E. (2022). Dexmedetomidine exerts an anti-inflammatory effect via α2 adrenoceptors to alleviate cognitive dysfunction in 5xFAD mice. Frontiers in Aging Neuroscience, 14, 978768.

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Vitamin C Modulates Cellular Responses

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Vitamin C (purity: > 99.9%) was purchased from Shandong Xinhua pharmaceutical CO., ltd. DMEM Medium, Fetal Bovine Serum (FBS), Wright-Giemsa staining, and Albumin Bovine V were purchased from MACGENE. RBC lysis Buffer, Sulforhodamine B (SRB), RNase A, Propidium Iodide (PI), and rat IgG were purchased from Solarbio. Triton X-100 solution, Ca²⁺ specific fluorescent probe Fluo-4/AM, JC-1 Staining Kit, One Step TUNEL Apoptosis Assay Kit, and ATP Determination Kit were purchased from Beyotime. FITC Rat Anti-Mouse CD11b was purchased from BD Biosciences. APC Rat Anti-Mouse CD206 was purchased from Miltenyi Biotec. Western blot Antibody Diluent was purchased from Epizyme. Western Bright™ ECL and Western Bright™ Peroxide were purchased from Advansta. Goat Anti-Rabbit IgG (H+L) HRP was purchased from Affinity Biosciences. Vimentin (5G3F10) Mouse mAb, Cleaved Caspase-3 (Asp175) Antibody (Alexa Fluor^® 488 Conjugate) and β-actin (13E5) Rabbit mAb were purchased from Cell Signaling Technology. E-cadherin Rabbit PolyAb was purchased from Proteintech.

Xu Y., Guo X., Wang G, & Zhou C. (2020). Vitamin C Inhibits Metastasis of Peritoneal Tumors By Preventing Spheroid Formation in ID8 Murine Epithelial Peritoneal Cancer Model. Frontiers in Pharmacology, 11, 645.

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Immunoblot Analysis of Protein Samples

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Proteins samples separated by 15% SDS-PAGE and blotted to a PVDF membrane. Membranes were blocked for 1 h with 5% not-fat milk in TBST buffer, and probed 1 hour at 4°C either in TBST with a 1:2000 anti-Flag antibody (ZEN-BIOSCIENCE, Catalogue No. 700002), or with a 1:1000 anti-His tag antibody (ZEN-BIOSCIENCE, Catalogue No. 251784). After washing by TBST buffer three times, membranes were incubated for 1 h with HRP-conjugated secondary antibodies (1:10000, Goat Anti-Rabbit IgG H&L (HRP), ZEN-BIOSCIENCE, Catalogue No. 511203) in TBST buffer. Chemiluminescence substrate (Abbkine, Catalogue No. K22020) was added and signal quantifications were done in BioRad imager. As for the acetylation assay, the PVDF membranes were firstly probed by Acetylated Lysine Antibody (Affinity, Catalogue No. DF7729), then incubated with Goat Anti-Rabbit IgG (H + L) HRP (Affinity, Catalogue No S0001).

Song Y., Zhang S., Ye Z., Song Y., Chen L., Tong A., He Y, & Bao R. (2022). The novel type II toxin–antitoxin PacTA modulates Pseudomonas aeruginosa iron homeostasis by obstructing the DNA-binding activity of Fur. Nucleic Acids Research, 50(18), 10586-10600.

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Immunohistochemical Analysis of Caspase-1 in Nucleus Pulposus

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The paraffin-embedded nucleus pulposus tissues were processed for immunohistochemical staining as previously described.^{33 (link)} The primary antibody used was caspase-1 (1:150, Proteintech, Chicago, USA, Cat. No.: 22915-1-AP) and the secondary antibody was goat anti-rabbit IgG (H+L) HRP (1:200, Affinity Biosciences, USA, Cat. #: S0001). The sections were visualized using a microscope (CTR4000B, Leica). Experiments were repeated three times independently.

Bai Z., Shou Z., Hu K., Yu J., Meng H, & Chen C. (2023). Melatonin protects human nucleus pulposus cells from pyroptosis by regulating Nrf2 via melatonin membrane receptors. Bone & Joint Research, 12(3), 202-211.

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Evaluating Angiogenic and Osteogenic Factors

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VEGFA and osteogenesis-related molecule expression were assessed by immunohistochemistry (IHC) using antibodies against VEGFA, RUNX2, and ALP on formalin-fixed paraffin-embedded (FFPE) tissue sections. The embedded samples were cut into 5 μm sections, deparaffinized with xylene, rehydrated using graded ethanol, antigen-retrieved, and then incubated with specific primary antibodies at 4°C overnight, and biotinylated secondary antibodies were incubated at room temperature for one hour. Then, sections were visualized with DAB precipitate and counterstained with hematoxylin. Images were captured under a light microscope (Nikon Ni-U, Japan). The mean density of protein expression was evaluated using the ImageJ software (USA).
The primary antibodies were anti-VEGFA (1 : 100, Abclonal, A12303), anti-RUNX2 (1 : 100, Affinity, AF5186), and anti-ALP (1 : 100, ImmunoWay, YT5563). The secondary antibodies were Goat Anti-Rabbit IgG (H + L) HRP (1 : 200, Affinity, S0001).

Wang Y., Luan S., Yuan Z., Wang S., Fan S., Ma C, & Wu S. (2022). The Combined Use of Platelet-Rich Plasma Clot Releasate and Allogeneic Human Umbilical Cord Mesenchymal Stem Cells Rescue Glucocorticoid-Induced Osteonecrosis of the Femoral Head. Stem Cells International, 2022, 7432665.

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