Rabbit pab

Manufactured by Abcam

Sourced in United States

Rabbit pAb is a primary antibody product developed in rabbits. It is designed for use in various immunodetection techniques, such as Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA).

Automatically generated - may contain errors

Lab products found in correlation

Acrylamide, by Bio-Rad (2 mentions) Amicon ultra column, by Merck Group (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Anti timp 1, by Cell Signaling Technology (1 mentions) Rabbit pab, by Cell Signaling Technology (1 mentions) Ti u fluorescent microscope, by Nikon (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Cd169 3d6 1.12, by Bio-Rad (1 mentions) B220 ra3 6b2, by Abcam (1 mentions) Prolong gold plus dapi, by Thermo Fisher Scientific (1 mentions) Igd 11 26c, by Southern Biotech (1 mentions) Tcs sp5 confocal microscope, by Leica (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Amersham ecl western blotting substrate, by GE Healthcare (1 mentions) Appropriate fluorescent secondary antibodies, by Thermo Fisher Scientific (1 mentions)

6 protocols using rabbit pab

BPIFB4 Expression and Secretion in HEK293T Cells

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HEK293T cells were grown in culture medium (DMEM, 10 % fetal bovine serum, 2 mM L-glutamine, 100 U penicillin/0.1 mg/ml streptomycin) and transfected with pRK5 vector encoding BPIFB4, or with an empty plasmid, using Lipofectamine 2000 (Life Technologies) according to the manufacturer’s protocol, in triplicates. 24 h after transfection, the medium is been changed and finally collect 48 h after the change. Protein content of medium is been concentrated using Amicon Ultra columns (Merck Millipore) and analyzed by Western blot. Protein were separated on 10 % SDS-PAGE at 100 V for 1 h or on 4–12 % SDS-PAGE at 100 V for 2 h and then transferred to a nitrocellulose or PVDF membrane. The membranes were incubated overnight with primary antibodies of anti-BPIFB4 (Abcam, rabbit pAb, 1:200) and anti-kallikrein 1 (Abcam, mouse pAb, 1:1000). After a triple wash, membranes were incubated for 1 or 2 h with the secondary antibody (Amersham Life Science, horseradish peroxidase-linked anti-rabbit IgG or anti-mouse IgG, 1:3000). The membranes were then washed four times and specific protein bands were detected with ECL Prime chemiluminescent agents (Amersham Life Science).

Villa F., Malovini A., Carrizzo A., Spinelli C.C., Ferrario A., Maciąg A., Madonna M., Bellazzi R., Milanesi L., Vecchione C, & Puca A.A. (2015). Serum BPIFB4 levels classify health status in long-living individuals. Immunity & Ageing : I & A, 12, 27.

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Western Blot Analysis of Cellular Proteins

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Proteins from whole cell lysates were separated by SDS-polyacrylamide gel electrophoresis (PAGE) with NuPAGE Novex Bis-Tris precast 4 to 12% gradient gels (Invitrogen, Carlsbad, CA, USA) and transferred to nitrocellulose membranes. The primary antibodies anti-actin (Mouse mAB; Sigma Life Science and Biochemicals), anti-TIMP1 (Rabbit pAB; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-CoL1A1 (Rabbit pAB;Thermo Fisher, Rockford, IL, USA), phosphorylated and unphosphorylated NFκB (RelA) (Rabbit pAB; Cell Signaling Technology, Inc., Danvers, MA, USA), phosphorylated NRF2 (Rabbit mAB; abcam, Cambridge, MA, USA), unphosphorylated NRF2 (Rabbit pAB; abcam), and phosphorylated and unphosphorylated SMAD3 (Rabbit mAB; Cell Signaling Technology, Inc.) were incubated overnight at 4°C. The secondary antibody, anti-mouse IgG (Donkey mAB; amersham Biosciences, Piscataway, NJ, USA), and anti-rabbit IgG (Sheep mAB; amersham Biosciences) were incubated for 60 minutes at room temperature.

Salloum S., Holmes J.A., Jindal R., Bale S.S., Brisac C., Alatrakchi N., Lidofsky A., Kruger A., Fusco D.N., Luther J., Schaefer E., Lin W., Yarmush M.L, & Chung R.T. (2016). HIV/HCV in hepatic and stellate cell lines reveals cooperative profibrotic transcriptional activation between viruses and cell types. Hepatology (Baltimore, Md.), 64(6), 1951-1968.

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Immunostaining of Cultured Neurons

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Cultured primary neurons were fixed in cold acetone/methanol for 10 minutes. After permeabilization and blocking with 0.1% Triton X100 in PBS containing 1% BSA for 1 hour, the neurons were then incubated with primary antibody against CAPON (rabbit pAb, 1:200; Abcam) and tyrosine hydroxylase (mouse mAb, 1:200; Sigma) in 1% BSA overnight at 4°C. Signals were visualized with antirabbit antibody conjugated to Alexa-488 (1:1000; Molecular Probes) and antimouse antibody conjugated to Alexa-594 (1:1000; Molecular Probes). Nuclear staining was performed with 4′,6-diamidino-2-phenylindole (DAPI, 1:1000; Sigma). Imaging was performed on a Nikon Ti-U fluorescent microscope.

Lu C.J., Hao G., Nikiforova N., Larsen H.E., Liu K., Crabtree M.J., Li D., Herring N, & Paterson D.J. (2015). CAPON Modulates Neuronal Calcium Handling and Cardiac Sympathetic Neurotransmission During Dysautonomia in Hypertension. Hypertension, 65(6), 1288-1297.

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Multicolor Immunofluorescence Analysis

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Organs were fixed in 1% formaldehyde–10 mM sodium periodate–75 mM l-lysine (24 h; 4°C), equilibrated in 30% sucrose (18 h; 4°C), and then frozen in optimal cutting temperature (OCT) matrix compound. Nine-micrometer sections were cut and then air dried (1 h; 23°C), blocked with 0.3% Triton X-100–5% normal goat serum (1 h; 23°C), and incubated (18 h; 4°C) with antibodies to enhanced green fluorescent protein (eGFP) (rabbit pAb; Abcam), macrophage receptor with collagenous structure (MARCO) (ED31; Serotec), IgM (biotin-conjugated goat pAb; Southern Biotech), IgD (11-26c; Southern Biotech), B220 (RA3-6B2; Abcam), and CD169 (3D6.112; Serotec).
GC B cell staining was with fluorescein-conjugated peanut agglutinin (PNA) (BD Biosciences). Sections were washed 3 times in PBS, incubated (1 h; 23°C) with Alexa 633-conjugated goat anti-rat IgG pAb, streptavidin-conjugated Alexa 568, and Alexa 488- or 568-conjugated goat anti-rabbit IgG pAb (Invitrogen), washed 3 times in PBS, and mounted in Prolong Gold plus DAPI (4′,6-diamidino-2-phenylindole) (Invitrogen). Fluorescence was visualized with a Leica TCS SP5 confocal microscope and analyzed with ImageJ.

Frederico B., May J.S., Efstathiou S, & Stevenson P.G. (2014). BAFF Receptor Deficiency Limits Gammaherpesvirus Infection. Journal of Virology, 88(8), 3965-3975.

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Western Blot Analysis of Cellular and Viral Proteins

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HAP1 cells were trypsinized and collected by centrifugation at 350 rcf for 10 min. The pellet was resuspended in Laemmli sample buffer containing 100 mM DTT, boiled for 5 min at 95°C and subjected to electrophoresis in 10% acrylamide (Bio-Rad) gels. Viruses were purified and concentrated over a 20% sucrose cushion (in TN buffer) at 110,000 rcf. Pelleted virus was resuspended in TN buffer overnight on ice. After addition of Laemmli sample buffer (1× final concentration, 100 mM DTT), samples were boiled for 5 min at 95°C and subjected to electrophoresis in 7% acrylamide (Bio-Rad) gels. Upon transfer to a nitrocellulose membrane (Millipore), the presence of cellular and viral proteins was probed with antibodies against GM130 (rabbit pAb, Abcam), FLAG (HRP-labeled mouse anti-FLAG mAb, Sigma) or the S2 subunit of MHV A59 [105] (link) (mouse anti-S2 mAb) diluted 1∶1000. When necessary, the blots were subsequently incubated with HRP-labeled rabbit anti-mouse or swine anti-rabbit antibodies (both diluted 1∶5000; DAKO). Binding of HRP-labeled antibodies was visualized using Amersham ECL Western blotting substrate (GE Healthcare Life Sciences) according to the manufacturer's instructions.

Burkard C., Verheije M.H., Wicht O., van Kasteren S.I., van Kuppeveld F.J., Haagmans B.L., Pelkmans L., Rottier P.J., Bosch B.J, & de Haan C.A. (2014). Coronavirus Cell Entry Occurs through the Endo-/Lysosomal Pathway in a Proteolysis-Dependent Manner. PLoS Pathogens, 10(11), e1004502.

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Multiplex Imaging of Lymph Nodes

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Six micron sections of frozen inguinal lymph nodes were thaw mounted onto glass slides and fixed in 4% paraformaldehyde for 30 min. After blocking with hydrogen peroxide, sections were stained for IL-2 and CD4 RNA using multiplexed fluorescent in situ hybridization (ACDbio). Following in situ hybridization, sections were incubated overnight with antibodies to FoxP3 (clone 236A/E7, Invitrogen) and either CD20 (Rabbit pAb, Abcam) or Ki67 (Rat clone OT1567, Abcam) on adjacent sections. Appropriate fluorescent secondary antibodies (Invitrogen) were added and all sections were counterstained with DAPI. Whole sections were imaged on an Aperio slide scanning system (Leica) at 40X.

Ollerton M.T., Folkvord J.M., La Mantia A., Parry D.A., Meditz A.L., McCarter M.D., D’Aquila R.T, & Connick E. (2022). Follicular regulatory T cells eliminate HIV-1-infected follicular helper T cells in an IL-2 concentration dependent manner. Frontiers in Immunology, 13, 878273.

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