Rabbit polyclonal igg

Manufactured by Abcam

Sourced in United Kingdom, United States

Rabbit polyclonal IgG is an antibody generated in rabbits through immunization with a specific antigen. This antibody recognizes and binds to multiple epitopes on the target antigen, providing a diverse set of binding interactions. Polyclonal antibodies are commonly used in various laboratory applications, such as Western blotting, ELISA, and immunohistochemistry, to detect and quantify the presence of target proteins in samples.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Triton x 100, by Merck Group (2 mentions) Mmp14, by Abcam (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Dp50 camera, by Olympus (1 mentions) Entellan, by Merck Group (1 mentions) Bx51 microscope, by Olympus (1 mentions) Cell f software, by Olympus (1 mentions) Microsystems microscope, by Leica (1 mentions) Hematoxylin, by Merck Group (1 mentions) Cy3 conjugated goat anti rabbit igg, by Dianova (1 mentions) 4 6 diamidino 2 phenylindole dihydrochloride dapi, by Carl Roth (1 mentions) Zen software, by Zeiss (1 mentions) Mowiol, by Leica (1 mentions) Fluorescence microscopy, by Leica (1 mentions) Phalloidin tritc, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) Texas red, by Abcam (1 mentions) Mouse monoclonal igg, by Elabscience (1 mentions) Goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Sheep anti-rabbit polyclonal IgG labeled with horseradish peroxidase Goat Polyclonal Anti-Rabbit IgG H&L (Alexa Fluor® 488), Polyclonal donkey anti-rabbit IgG-HRP Goat anti-rabbit IgG polyclonal antibody HRP-conjugated goat anti-rabbit IgG polyclonal antibody Goat polyclonal antibody to rabbit IgG Goat polyclonal anti-rabbit IgG-HRP Rabbit anti-sheep IgG H&L (FITC) polyclonal antibody AF594-conjugated donkey IgG F(ab’)2 fragment polyclonal antibody to rabbit IgG Polyclonal rabbit anti-mouse serum albumin IgG primary antibody

20 protocols using rabbit polyclonal igg

Immunofluorescence Staining of Pancreatic Tissues and Pancreatic Stellate Cells

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Immunofluorescence staining for pancreatic tissues was performed as described previously.^{30 (link)} Briefly, paraffin-embedded pancreas tissue sections (4 μm) were deparaffinized and rehydrated in a graded series of ethanol. After blocking with 2% normal BSA, sections were incubated with mouse anti-rat glial fibrillary acidic protein (GFAP, to stain quiescent PSCs) or α-SMA antibody (1:100) and rabbit anti-rat Grp78 (1:100) overnight at 4°C. After washing, slides were incubated with anti-rabbit Alexa 488-conjugated IgG antibody and anti-mouse Alexa 555-conjugated IgG antibody for 1 hour.
Immunofluorescence staining for PSCs was performed as described previously.^{30 (link)} Briefly, PSCs were serum starved, incubated for 24 hour at 37°C, and fixed in 4% paraformaldehyde. Double-immunofluorescence staining for α-SMA/Grp78 was performed in a similar manner as that described above. Nuclear counterstaining was performed using Hoechst 33342 (Invitrogen). The fluorescence of the slides was analyzed under a confocal laser scanning microscope (Nikon A1/C1, Tokyo, Japan). Sections without primary antibody or with polyclonal rabbit IgG (Abcam) were included in each staining experiment as negative controls (data not shown).

Lee L., Ito T., Nakamura T., Jensen R.T., Igarashi H, & Takayanagi R. (2017). Anti-fibrotic Effect of Saturated Fatty Acids via Endoplasmic Reticulum Stress Response in Rat Pancreatic Stellate Cells. Pancreas, 46(3), 385-394.

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Megalin Immunohistochemistry in OSCC

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Immunohistochemical staining of megalin protein was conducted using DAKO EnVision + System, Peroxidase (DAB) kit (DAKO Cytomation, Santa Clara, CA, USA) on 4 µm thick serial sections of paraffin-embedded OSCC tissue. In brief, after being deparaffinized and rehydrated, tissues underwent heat-mediated epitope retrieval by microwave heating in a 10-mM citrate buffer of pH = 6.0. Having subsequently been treated with blocking solution, tissue slides were incubated with polyclonal rabbit anti-megalin IgG (H-245, Santa Cruz Biotechnology, Dallas, TX, USA; diluted 1:100 in 1% BSA in PBS) over 12 h in a humid chamber at 4 °C. As a secondary antibody, a peroxidase-labeled polymer linked to goat anti-rabbit antibodies was applied for 30 min at room temperature. Immunoreactions were visualized by 3,3′-Diaminobenzidine (DAB) and sections were counterstained with hematoxylin. Following dehydration, slides were covered with Entellan (Sigma-Aldrich, Hamburg, Germany) and analyzed by an Olympus BX51 microscope equipped with a DP50 camera and Cell^F software (Olympus, Tokyo, Japan). The specificity of antibody binding was verified performing negative controls by substitution of the megalin antibody with an isotype-matched control antibody (polyclonal rabbit IgG, Abcam, Cambridge, UK) applied in the same conditions. Negative control slides did not show immunohistochemical signals.

Zulijani A., Milardović A., Kovač Z., Perić B, & Jakovac H. (2023). Megalin Expression in Primary Oral Squamous Cell Carcinoma Is Associated with the Presence of Lymph Node Metastases, Vascular Invasion, and Lower Overall Survival. Current Issues in Molecular Biology, 45(4), 2757-2766.

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Immunohistochemical Staining of Glioma Tissues

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First, paraffin-embedded sections (2 µm-thick) of glioma tissue of various grades were prepared using conventional dehydration and paraffin embedding procedures. In brief, paraffin sections were baked in citrate buffer (1 h, 65°C), washed with water, and subjected to high-pressure antigen retrieval. Endogenous peroxidase activity was blocked by incubation for 10 min with 3% H₂O₂. After blocking, the sections were incubated with an anti-A3C antibody (1:100; 10591-1-AP, Proteintech, USA) at 37°C for 1 h. The sections were washed 5 times with PBS and were then incubated with a secondary antibody (polyclonal rabbit IgG, Abcam). After staining with diaminobenzidine (DAB), the sections were washed with PBS to remove excess stain, counterstained with hematoxylin, differentiated with hydrochloric acid alcohol, and blued with tap water. Finally, the sections were dehydrated through an alcohol gradient, mounted in neutral resin in xylene, and observed under an M2000 LED microscope (Leica, Germany). Images were processed using Motic DSAssistant software and staining was quantified by measurement of the optical density using ImageJ software (v1.46, Bethesda, MD, USA).

Zhang S., Guo Y., Hu Y., Gao X., Bai F., Ding Q., Hou K., Wang Z., Sun X., Zhao H., Qu Z, & Xu Q. (2023). The role of APOBEC3C in modulating the tumor microenvironment and stemness properties of glioma: evidence from pancancer analysis. Frontiers in Immunology, 14, 1242972.

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Histomorphometric Analysis of Bone Tissue

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Paraffin-embedded femurs were sectioned (5 μm), deparaffinized, and then stained with hematoxylin and eosin (H&E) for morphological analysis. For immunohistochemistry staining, deparaffinized and dehydrated tissue sections were re-hydrated before being subjected to antigen retrieval, and then blocked with diluted normal serum for at least 1–1.5 h at 25 °C to eliminate nonspecific binding. The slides were incubated with primary antibodies against osteocalcin (OCN) (1:300; Abcam) overnight at 4 °C. After careful washing, the sections were incubated with horseradish peroxidase conjugates to detect positive signals, followed by counterstaining with hematoxylin (Sigma-Aldrich). Slides incubated with polyclonal rabbit IgG (Abcam) served as negative controls. Pictures were captured and monitored using a Leica Microsystems microscope (Leica Microsystems Ltd.), while the ImageJ software (National Institutes of Health) was used to analyze the number or area of adipocytes and osteoblasts.

Chen G., Zhuo Y., Tao B., Liu Q., Shang W., Li Y., Wang Y., Li Y., Zhang L., Fang Y., Zhang X., Fang Z, & Yu Y. (2020). Moderate SMFs attenuate bone loss in mice by promoting directional osteogenic differentiation of BMSCs. Stem Cell Research & Therapy, 11, 487.

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Immunofluorescent Detection of POR Protein

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Cells growing on glass slides were fixed for 30 min in 5% formaldehyde solution, followed by 10 min permeabilisation in 0.2% Triton-X100 solution. Blocking of unspecific binding sites occurred for 30 min in 3% BSA solved in PBS. All steps were performed at ambient temperature. Visual detection of POR protein expression was realised using polyclonal rabbit-anti-POR IgG as primary antibody (1 mg/mL, Abcam) at 1:100 dilution and a Cy3 conjugated goat-anti-rabbit IgG (1:200, Dianova GmbH, Hamburg, Germany) as secondary antibody. Incubation of the samples with the primary antibody occurred over night at 4°C and with the secondary antibody for 1 hour at ambient temperature. For assessment of background binding, samples were stained with a primary antibody of the same isotype (polyclonal rabbit IgG, 1 mg/mL, Abcam) but without target binding properties serving as isotype control. Staining of genomic DNA was realised by 5 min incubation of the samples with 4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI, 0.2 μg/mL in PBS, Carl Roth GmbH + Co. KG, Karlsruhe, Germany). High resolution pictures were taken by using a LSM800 confocal Laser Scanning Microscope system and ZEN software for picture post processing (Carl Zeiss Microscopy GmbH, Jena, Germany).

Schulz C., Kammerer S, & Küpper J.H. (2019). NADPH-cytochrome P450 reductase expression and enzymatic activity in primary-like human hepatocytes and HepG2 cells for in vitro biotransformation studies. Clinical Hemorheology and Microcirculation, 73(1), 249-260.

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Immunohistochemical Analysis of Inflammatory Cells in Takayasu Arteritis

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Detection of IL-33 1 , ST2 1 , and tryptase 1 cells was performed on fixed, paraffin-embedded samples (aorta) from patients with TAK and noninflammatory aorta controls. After dewaxing in baths of xylene and ethanol, slides were submitted to antigen retrieval by heating (1108C) in citrate buffer at pH 6.0. Before incubation with primary antibodies, Fc receptors were blocked with normal goat serum (3.3%). Slides were then incubated overnight with mouse monoclonal anti-IL-33 (dilution 1:1000; Enzo Life Sciences, Farmingdale, NY), rabbit polyclonal anti-ST2 (dilution 1:100; Sigma-Aldrich), monoclonal mouse anti-tryptase (1/200; Dako, Agilent, Santa Clara, Calif), or with isotype control: polyclonal rabbit IgG or monoclonal mouse IgG (Abcam, Cambridge, UK). Slides were then incubated for 2 hours at room temperature with Cy3conjugated goat anti-mouse immunoglobulin (working dilution 1:1000; Jackson ImmunoResearch, West Grove, Pa) and Alexa 488 donkey anti-rabbit Ig (working dilution 1:1000; Life Technologies, Thermo Fisher Scientific), mounted in Mowiol and evaluated under fluorescence microscopy (Leica Microsystems, Wetzlar, Germany). MCs were also quantified by counting the number of tryptase 1 cells per field in 100 fields per sample.

Le Joncour A., Desbois A.C., Leroyer A.S., Tellier E., Régnier P., Maciejewski-Duval A., Comarmond C., Barete S., Arock M., Bruneval P., Launay J.M., Fouret P., Blank U., Rosenzwajg M., Klatzmann D., Jarraya M., Chiche L., Koskas F., Cacoub P., Kaplanski G, & Saadoun D. (2022). Mast cells drive pathologic vascular lesions in Takayasu arteritis. The Journal of allergy and clinical immunology, 149(1).

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Immunofluorescence Staining of vWF and CD41

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After dewaxing, rehydration, antigen retrieval with citric acid (pH6) at 95 °C for 10 min, and blocking with 10% goat serum, slides were incubated with primary antibodies (1:500 Rabbit polyclonal IgG against vWF, Sigma, Gillingham UK; 1:25 Mouse monoclonal IgG1 against CD41 platelet marker integrin αIIb, clone B-9, Santa Cruz, CA, USA) at 4 °C overnight. An hour-long incubation of slides with 1% Bovine serum albumin/Tris-buffered saline (TBS) suspended 4′,6-diamidino-2-phenylindole (DAPI) (1:100 nuclei stain, Sigma) and secondary antibodies (conjugated with Cyanide Dyes, Cy2 1:100, Cy3 1:500, Jackson ImmunoResearch, Ely, UK) was completed at room temperature. IgG isotype control (1:400 Rabbit polyclonal IgG, Abcam, Cambridge, UK) was used to replace primary antibody against vWF to ensure specific binding. For assessment of nonspecific binding of secondary antibodies, reagent control was carried out by omitting all primary antibodies.

Cheng W.C., Wilkie L., Kurosawa T.A., Dobromylskyj M., Priestnall S.L., Luis Fuentes V, & Connolly D.J. (2021). Immunohistological Evaluation of Von Willebrand Factor in the Left Atrial Endocardium and Atrial Thrombi from Cats with Cardiomyopathy. Animals : an Open Access Journal from MDPI, 11(5), 1240.

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Cytoskeletal Dynamics in HK-2 Cells

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HK-2 cells cultured under static condition were seeded in confocal culture dishes at a seeding density of 2 × 10⁵/cm² . Cells cultured under flow condition were as described in Section 2.3. For filamentous actin (F-actin) staining, cells were fixed in 4% paraformaldehyde for 20 min, rinsed three times with PBS, permeabilized with 0.1% Triton X-100 (Sigma–Aldrich) for 10 min, and then rinsed three times with PBS. Next, cells were blocked with 1% BSA at room temperature and incubated with 1:20 diluted TRITC-phalloidin (Invitrogen) at 37 °C for 30 min. Then, the cells were rinsed three times with PBS and stained with 4′,6-diamidino-2-phenylindole (DAPI) (Beyotime Biotechnology, Chengdu, China) for 10 min. The fluorescence images were taken by confocal microscopy.
For immunofluorescence staining of megalin and clathrin, cells were fixed as described above and incubated in 1% BSA at room temperature for 1 h. After rinsing with PBS three times, primary antibodies for megalin or clathrin (rabbit polyclonal IgG, Abcam) were added and incubated at 4 °C overnight. Next, the secondary antibody (goat anti-rabbit IgG/Alexa fluor 488 or FITC, Zhongshanjinqiao Biotechnology, Beijing, China) was added and incubated at room temperature for 1 h. The fluorescence images were taken by confocal microscopy.

Xu Y., Qin S., Niu Y., Gong T., Zhang Z, & Fu Y. (2019). Effect of fluid shear stress on the internalization of kidney-targeted delivery systems in renal tubular epithelial cells. Acta Pharmaceutica Sinica. B, 10(4), 680-692.

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Characterization of Decellularized Testicular Scaffolds

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Fixation of the scaffolds was performed by incubation
in 10% formalin solution in PBS at 25°C for 24-48 hours.
The fixed scaffolds were then dehydrated by incubation
in graded alcohol (each alcohol for 20 minutes). After
embedding them in paraffin, they were cut into 5 µm-thick
sections for histological evaluation. Hstaining was performed on paraffin sections for evaluation
of detergent efficacy to remove the cells and debris from
the testes. Preservation of glycosaminoglycans (GAGs)
and collagen in decellularised scaffolds were assessed by
alcian blue (Sigma, USA) and Masson’s trichrome staining
(Sigma, USA), respectively. Alcian blue was diluted 1:100
in hydrochloric acid (0.1 M). On these samples nuclear
fast red (Sigma, USA) was used for counter-staining
(12 (link), 13 (link)). Also, preservation of ECM proteins, including
fibronectin, collagen IV, and laminin in decellularised
scaffolds was evaluated by immunohistochemistry
(IHC). Initially, the sections were transferred to a 60°C
oven for de-waxing, then further cleared in xylene. Later,
they were rehydrated by alcohol gradient and washing
in water. Then they were incubated in citrate (10 mM
pH=6.0) for 20 min for antigen retrieval. Then the samples
were permeabilized by triton X-100 for 40 minutes and
incubated with anti-fibronectin (mouse monoclonal IgG,
Elabscience Biotechnology Inc., USA), anti-collagen IV
(mouse monoclonal IgG, Elabscience Biotechnology Inc.,
USA), and anti-laminin (rabbit polyclonal IgG, Abcam,
USA). The secondary antibody was Alexa Fluor 488 (goat
anti-mouse IgG, Invitrogen, USA) and Texas Red (Goat
anti-rabbit IgG, Abcam, USA). Photomicrographs were
taken with an Olympus microscope (Olympus, Center
Valley, PA, USA).

Majidi Gharenaz N., Movahedin M, & Mazaheri Z. (2019). Three-Dimensional Culture of Mouse Spermatogonial Stem Cells Using A Decellularised Testicular Scaffold. Cell Journal (Yakhteh), 21(4), 410-418.

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In vivo Bladder Immunostaining Protocol

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For in vivo bladder section, immunostaining was performed according to published methods [29 (link),30 (link),31 (link)]. The bladder whole mounts were fixed in cold 4% paraformaldehyde in 0.1 M PBS overnight, washed with PBS, blocked with 10% NGS in PBS/0.5% Triton X-100 for 1hr at room temperature (RT), and then incubated with the primary antibody to laminin (Abcam, rabbit polyclonal IgG, 1:100 catalog no. ab11575), α-SMA (Abcam, rabbit monoclonal IgG, 1:100, catalog no. ab5694) and LC3 (Santa Cruz, mouse monoclonal IgG, 1:50, catalog no. sc-376404) at 4 °C over two night. Tissues were washed in PBS/0.5% Triton X-100 15 min on a rocker and incubated with secondary antibodies at RT for 2 h. After washing, DAPI was added, and the whole mounts were transferred to a glass slide and coverslipped with a 100 mm adhesive spacer in Prolong Gold anti-fade reagent (Invitrogen). Double staining of the green-stained laminin and the red-stained LC3 in the bladder whole mount were captured using confocal laser scanning microscopy. In each experiment, negative controls without the primary antibody were performed to elucidate non-specific immunostaining.

Lu J.H., Wu Y.H., Juan T.J., Lin H.Y., Lin R.J., Chueh K.S., Lee Y.C., Chang C.Y, & Juan Y.S. (2021). Autophagy Alters Bladder Angiogenesis and Improves Bladder Hyperactivity in the Pathogenesis of Ketamine-Induced Cystitis in a Rat Model. Biology, 10(6), 488.

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