Polyclonal rabbit antibodies

Manufactured by Abcam

Polyclonal rabbit antibodies are primary antibodies produced by immunizing rabbits with a specific antigen. These antibodies recognize multiple epitopes on the target antigen, providing a broader specificity and increased sensitivity in immunoassays and other applications.

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Lab products found in correlation

Panreac blocking buffer, by AppliChem (1 mentions) Goat anti rabbit igg horseradish peroxidase hrp, by Bio-Rad (1 mentions) Phosphatase inhibitor, by Merck Group (1 mentions) Protease inhibitor, by Roche (1 mentions) Metamorph premier image analysis software, by Molecular Devices (1 mentions) Fulvestrant, by Bio-Techne (1 mentions) Chenodeoxycholic acid, by Merck Group (1 mentions) Anti bsp, by Santa Cruz Biotechnology (1 mentions) Rabbit polyclonal antibodies against, by Santa Cruz Biotechnology (1 mentions) 17β estradiol, by Merck Group (1 mentions) 4 hydroxytamoxifen, by Merck Group (1 mentions) Virtual slide system vs110, by Olympus (1 mentions)

4 protocols using polyclonal rabbit antibodies

Western Blot Analysis of P2X Receptors

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Total lysates were prepared by homogenizing various tissues in RIPA buffer supplemented with protease inhibitors (Roche) and phosphatase inhibitors (Sigma). Protein samples (25 μg per lane) were separated by SDS-PAGE. Blots were blocked for 2 h in PanReac Blocking buffer (AppliChem), incubated for 1 h with appropriate primary antibody and 2 h with HRP-conjugated secondary antibody. Polyclonal rabbit antibodies directed against the C-terminal peptides of P2X4 and P2X7 were from Abcam (Cat. No. ab243734, Cat. No. ab229453). The P2X7 antibody detected two bands at around 75 kDa in BAT of Balb/c mice. The upper band was also detectable in knockout mice, indicating that as a non-specific cross-reactivity of the antibody. P2X5 rabbit polyclonal antibody was purchased from Thermo Fisher (Cat. No. PA5-41079) and the loading control γ-tubulin rabbit monoclonal antibody from Abcam (Cat. No. ab179503). The secondary antibody, goat-anti-rabbit IgG horseradish peroxidase (HRP), was purchased from Bio-Rad. Detection was performed on Amersham Imager600 using luminol and para-hydroxycoumarinic acid–based chemiluminescence substrate.

Tian T., Heine M., Evangelakos I., Jaeckstein M.Y., Schaltenberg N., Stähler T., Koch-Nolte F., Kumari M, & Heeren J. (2020). The P2X7 ion channel is dispensable for energy and metabolic homeostasis of white and brown adipose tissues. Purinergic Signalling, 16(4), 529-542.

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Analyzing Lupus-Induced Diffuse Alveolar Hemorrhage

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Formalin-fixed, paraffin-embedded archived human lung biopsy tissue from a 19-year-old woman with lupus nephritis who developed massive hemoptysis and DAH was sectioned (4 μm) and stained with hematoxylin & eosin (H&E). Pristane-treated mice were euthanized at 14-d and lungs were formalin-fixed. DAH was evaluated by gross inspection of excised lungs and confirmed by microscopically. Tissue sections were subjected to antigen retrieval and analyzed by TUNEL (ApopTag Peroxidase In Situ Apoptosis Detection Kit, Chemicon/Millipore, Danvers, MA). Neutrophil elastase was detected by IHC with polyclonal rabbit antibodies (Abcam, Cambridge, MA, 1:50 dilution for 60-min) and quantified morphometrically. The expression area and staining intensity were quantified using MetaMorph Premier Image Analysis Software (Molecular Devices Corporation, Sunnyvale, CA). Staining intensity (thresholded area) was expressed as percentage of total examined lung cell area after subtracting noncellular space from total area.
Staining with oil red-O was performed on 10 μm frozen sections of lung tissue from pristane-treated mice or untreated controls [11 (link)]. Tissue was counterstained with Mayer’s hematoxylin and viewed microscopically.

Zhuang H., Han S., Lee P.Y., Khaybullin R., Shumyak S., Lu L., Chatha A., Afaneh A., Zhang Y., Xie C., Nacionales D., Moldawer L., Qi X., Yang L.J, & Reeves W.H. (2017). Pathogenesis of diffuse alveolar hemorrhage in murine lupus. Arthritis & rheumatology (Hoboken, N.J.), 69(6), 1280-1293.

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Osteogenic Differentiation Pathway Regulation

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17β-estradiol (E₂), 4-hydroxytamoxifen, chenodeoxycholic acid (CDCA) and lithocholic acid (LCA) were obtained from Sigma Aldrich®, fulvestrant (ICI 182,780) from Tocris® (Bristol, UK) and Z-guggulsterone from Enzo Life Sciences®. These compounds were first prepared as stock solution at 10^− 2 M in ethanol 99.8%, except for 4-hydroxytamoxifen and fulvestrant, which were prepared as stock solution at 10^− 3 M. All preparations were done under sterile conditions.
A murine monoclonal antibody against FXR was purchased from R&D Systems®. Polyclonal rabbit antibodies against osteopontin (anti-OPN) and RUNX2 were purchased from Abcam®. Polyclonal rabbit antibodies against estrogen receptor alpha, osteocalcin (anti-OC) and a mouse monoclonal antibody against bone sialoprotein (anti-BSP) came from Santa Cruz Biotechnologies®. Except for the RUNX2 antibody (dilution 1/100), all primary antibodies were diluted at 1/50 in PBS (pH 7.2) containing casein 0.05% (Sigma Aldricht®) for immunofluorescence staining.

Absil L., Journé F., Larsimont D., Body J.J., Tafforeau L, & Nonclercq D. (2020). Farnesoid X receptor as marker of osteotropism of breast cancers through its role in the osteomimetism of tumor cells. BMC Cancer, 20, 640.

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Quantitative Histopathological Analysis of Liver

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Formalin-fixed, paraffin-embedded livers were cut at 5 microns and stained with picrosirius red, and immunohistochemically for cytokeratin-19 (CK-19), CD3 (T-cells), and CD45R (B-cells) by the Investigative Histopathology Laboratory at Michigan State University as described previously (Joshi et al. 2015 (link); Joshi et al. 2016a (link)). Primary antibodies utilized were polyclonal rabbit antibodies (Abcam, Cambridge, MA) and detected by HRP-conjugated polymer detection systems. Images comprising the entire left lateral lobe (>500 images) were captured using a Virtual Slide System VS110 (Olympus, Hicksville, NY) with a 20× objective. The area of positive sirius red and CK-19 staining was determined in an automated and unbiased fashion using a batch macro and the color de-convolution tool in ImageJ. Positive area for each stain was expressed individually and as a ratio of sirius red/CK-19. Similarly, the number of CD3 and CD45R-positive cells per image was determined using a batch macro, the color de-convolution tool, and particle analysis function of ImageJ.

Joshi N., Kopec A.K., Cline-Fedewa H, & Luyendyk J.P. (2016). Lymphocytes contribute to biliary injury and fibrosis in experimental xenobiotic-induced cholestasis. Toxicology, 377, 73-80.

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