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Rabbit polyclonal antibodies

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Rabbit polyclonal antibodies are a type of antibody reagent produced by immunizing rabbits with a specific antigen. These antibodies are a heterogeneous mixture that recognize multiple epitopes on the target antigen. They are commonly used in various research and diagnostic applications.

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Lab products found in correlation

Polysulfoethyl column, by Nest Group (3 mentions) Ab6721, by Abcam (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Quantity one software, by Bio-Rad (1 mentions) Ecl plus detection system, by Thermo Fisher Scientific (1 mentions) Gel 3100 chemiluminescent and fluorescent imaging system, by Sagecreation (1 mentions) Smifh2, by Merck Group (1 mentions) Cytod, by Merck Group (1 mentions) Ck 666, by Merck Group (1 mentions) Hrp conjugated secondary antibody, by Abcam (1 mentions) Safranin o solution, by Merck Group (1 mentions) Hematoxylin solution, by Merck Group (1 mentions) Goat polyclonal antibodies, by Santa Cruz Biotechnology (1 mentions) Fibronectin, by Cell Signaling Technology (1 mentions) Glyceraldehyde 3 phosphate dehydrogenase, by Cell Signaling Technology (1 mentions) P src, by Cell Signaling Technology (1 mentions) Sybr green 1 master kit, by Roche (1 mentions) Short interfering rnas sirnas, by GenePharma (1 mentions)

21 protocols using rabbit polyclonal antibodies

1

Quantitative Western Blot Analysis

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Protein extracts from the same kidneys that were used for 2DE (30 μg protein) were separated by SDS-PAGE (n = 3/group). After electrophoresis proteins were transferred to a PVDF membrane (BioRad). Membranes were probed with rabbit polyclonal antibodies (Abcam) against triosephosphate, aminoacylase 1, rabbit polyclonal anti-MLC2, phosphoglycerate kinase 1 according to the supplier’s instructions. Goat anti-rabbit secondary antibodies, tagged with Alexa Fluor647 (ThermoFisher Scientific, Waltham, MA, USA), were used and membranes developed with VersaDoc5000 using appropriate filters. Band densities were determined with Quantity One software (BioRad). Actin was used as a protein loading control.
Moser M.A., Sawicka K., Sawicka J., Franczak A., Cohen A., Bil-Lula I, & Sawicki G. (2020). Protection of the transplant kidney during cold perfusion with doxycycline: proteomic analysis in a rat model. Proteome Science, 18, 3.
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2

Western Blotting Analysis of Cellular Stress Signaling

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Western blotting was performed as described previously (Zhu et al., 2012b (link)). The membranes were blocked for 1 h in Tris-buffered saline containing 0.05% Tween 20 and 5% nonfat milk and then probed for 1 h with the following primary antibodies: mouse monoclonal anti-Cap IgG (produced in our laboratory),
mouse monoclonal anti-Bcl-2 (Abcam, Cambridge, UK), rabbit monoclonal antibodies to GRP78, eukaryotic initiation factor 2α (eIF2α) phospho and total eIF2α (Abcam), rabbit polyclonal antibodies to PERK phospho, total PERK, CCAAT/enhancer-binding protein homologous protein (CHOP) (Abcam), goat anti-activating transcription factor 4 (ATF4) polyclonal antibody (Abcam), and mouse monoclonal antibodies to β-actin, Flag and GFP (MultiSciences, Hangzhou, China). All antibodies were found to react with target molecules from porcine cell lines used in this study. Blots were washed and then incubated for another hour with goat anti-rabbit, anti-mouse or donkey anti-goat horseradish peroxidase-labeled antibodies (KPL, Gaithersburg, MD, USA). The blots were revealed using the ECL Plus detection system under conditions recommended by the manufacturer (Thermo, Marina, CA, USA). Images were captured by the Gel 3100 Chemiluminescent and Fluorescent Imaging System (Sagecreation, Beijing, China).
Zhou Y.S., Gu Y.X., Qi B.Z., Zhang Y.K., Li X.L, & Fang W.H. (2017). Porcine circovirus type 2 capsid protein induces unfolded protein response with subsequent activation of apoptosis. Journal of Zhejiang University. Science. B, 18(4), 316-323.
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3

Immunoprecipitation of BAF53a and Tip60

Cited 1 time
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Western blotting was performed and immunoprecipitation according to our previous work [31 (link)]. For immunoprecipitation, rabbit polyclonal antibodies (Abcam) against BAF53a or Tip60 were used. The cytosolic fraction (2 mg/ml) from subcellular fractionation assay was used as input (IN). As negative control, no antibody was added to a same amount of IN and beads (Santa Cruz Biotechnology).
Messina G., Prozzillo Y., Monache F.D., Santopietro M.V, & Dimitri P. (2022). Evolutionary conserved relocation of chromatin remodeling complexes to the mitotic apparatus. BMC Biology, 20, 172.
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4

Microscopic Actin Cytoskeleton Analysis

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Mouse monoclonal antibodies to Siglec-1 (Hsn 7D2) and rabbit polyclonal antibodies against cofilin and phospho-cofilin (phospho S3) were obtained from Abcam. Rabbit antibodies against pERM (48G2) and pMLC were purchased from Cell Signaling. Anti-Mouse IgG F(ab) ATTO488 was from Hypermol and Anti-Rabbit IgGF(ab) Cy3 was from Jackson Immunoresearch. Donkey anti-mouse AlexaFluor Plus 488 secondary antibody was from Thermo Fisher. The SIR-Actin Spirochrome Kit (CY-SC001), the Rho Inhibitor I ADP ribosylation of Rho Asn-41 (CT04) and Rho Activator II (CN03) were obtained from Cytoskeleton. The phosphor-ezrin inhibitor NSC668394 was from Calbiochem. The pan-formin inhibitor SMIFH2, the Arp2/3 inhibitor CK-666 and the actin polymerization inhibitor CytoD were purchased from SIGMA. The ROCK inhibitor Y-27632 was obtained from Merk Millipore.
Gutiérrez-Martínez E., Benet Garrabé S., Mateos N., Erkizia I., Nieto-Garai J.A., Lorizate M., Borgman K.J., Manzo C., Campelo F., Izquierdo-Useros N., Martinez-Picado J, & Garcia-Parajo M.F. (2023). Actin-regulated Siglec-1 nanoclustering influences HIV-1 capture and virus-containing compartment formation in dendritic cells. eLife, 12, e78836.
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5

TanIIA Compound Characterization Protocol

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TanIIA (purity > 98%, NM_20100205) was purchased from the Zhejiang Institute of Drug Control (Zhejiang, China). RPMI-1640, fetal bovine serum (FBS), penicillin, pancreatic enzyme, and streptomycin were purchased from Hyclone (Logan, USA). All the rabbit polyclonal antibodies and HRP-conjugated secondary antibodies were purchased from Abcam (California, USA).
Wang J.F., Feng J.G., Han J., Zhang B.B, & Mao W.M. (2014). The Molecular Mechanisms of Tanshinone IIA on the Apoptosis and Arrest of Human Esophageal Carcinoma Cells. BioMed Research International, 2014, 582730.
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6

Quantitative Western Blot Analysis of SGLT1 and SGLT2

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For each electrophoresis lane, equal amounts of protein (30 μg) were mixed with loading buffer and boiled for five minutes. Samples were separated on an 8% sodium dodecyl sulfate-polyacrylamide gel and then transferred to polyvinylidene fluoride (PVDF) membrane. PVDF blots were blocked by incubation in PBST (20 mM PBS, pH 7.4, and 0.1% Tween 20) containing 5% nonfat skim milk overnight at 4 °C and then incubated with a rabbit polyclonal antibodies against SGLT1 (1: 1000 dilution), SGLT2 (1: 200 dilution), and β-actin (1: 3000 dilution) for 2 h at 37 °C (Abcam Inc., Cambridge, MA). After three washing steps (PBST, each step 5 min), a horseradish peroxidase (HRP)-conjugated polyclonal secondary anti-rabbit antibody (1: 20,000 dilution) was added and incubated for 1 h. After three washing steps, the bands were visualized with enhanced chemiluminescence detection kit (ECL) (Thermo Scientific, Waltham, MA). Image J software (NIH) was used for densitometric analysis of the bands.
Pourfarjam Y., Rezagholizadeh L., Nowrouzi A., Meysamie A., Ghaseminejad S., Ziamajidi N, & Norouzi D. (2016). Effect of Cichorium intybus L. seed extract on renal parameters in experimentally induced early and late diabetes type 2 in rats. Renal Failure, 39(1), 211-221.
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7

Visualizing Neocartilage Deposition and Distribution

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To visualize neocartilage deposition and distribution, immunofluorescence of collagen I, II, and X and histological staining of sGAG were performed on encapsulated chondrocytes after 3 weeks in culture. Cell-laden hydrogels were first fixed with 4% paraformaldehyde and dehydrated using 30% sucrose solution overnight before freezing in OCT compound (Tissue-Tek). Sections were prepared and stained for type I, II, and X collagen using rabbit polyclonal antibodies (Abcam) in accordance with a previously reported immunostaining protocol.24 (link) For sGAG histological staining, sections were prepared and stained as previously reported25 (link) using hematoxylin solution (Sigma-Aldrich) for nuclei staining and 0.1% safranin-O solution (Sigma) for staining sGAG.
Zhu D., Trinh P., Liu E, & Yang F. (2018). Biochemical and Mechanical Gradients Synergize To Enhance Cartilage Zonal Organization in 3D. ACS biomaterials science & engineering, 4(10), 3561-3569.
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8

Antibody Generation and Immunofluorescence Assay

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A LIMP-2 sheep polyclonal antibody was generated against the peptide sequence CKKLDDFVETGDIRTMVFP (Mimotopes Pty. Ltd., VIC, Australia). Rabbit polyclonal antibodies (Abcam PLC, Cambridge United Kingdom) were against Appl1 (0.4 μg/mL), Appl2 (0.4 μg/mL), Rab4 (1 μg/mL), TGN46 (10 μg/mL), TfR1 (1 μg/mL), TfR2 (1 μg/mL). Akt (1/1000) and phospho-Akt (Thr308 1/1000) from Cell Signalling Technology Inc., MA, USA, and HRP-conjugated anti-GAPDH (1/20000 Sigma Aldrich Pty. Ltd., NSW, Australia). Goat polyclonal antibodies (Santa Cruz Biotechnology, CA, USA) were against Rab5 (1 μg/mL), Rab7 (1 μg/mL) and EEA1 (1 μg/mL). A LAMP-1 (1 μg/mL) mouse monoclonal BB6 was provided by Professor Sven Carlsson (Umea University, Sweden). HRP-conjugated secondary antibodies for Western blot analysis included anti-goat/sheep (1/2000, Merck Millipore Pty. Ltd., VIC, Australia), anti-rabbit (1/2000) and anti-mouse (1/2000) (Sigma Aldrich). The secondary and other antibody conjugated fluorophores that were used included Alexa Fluor® 488 (1/250), Alexa Fluor® 633 (1/250), Transferrin-633 (1/1000), Phalloidin-488 (1/100), and LysoTracker® (5 μM); all from Life Technologies Pty. Ltd., VIC, Australia.
Johnson I.R., Parkinson-Lawrence E.J., Shandala T., Weigert R., Butler L.M, & Brooks D.A. (2014). Altered Endosome Biogenesis in Prostate Cancer has Biomarker Potential. Molecular cancer research : MCR, 12(12), 1851-1862.
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9

Molecular Profiling of Cell Signaling

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Phosphate-buffered saline (PBS), Roswell Park Memorial Institute medium (RPMI) 1640, and fetal bovine serum (FBS) purchased from Hyclone (Logan, UT, USA) were used. Rabbit polyclonal antibodies against human HERG1 and TXNDC5 were purchased from Abcam (Cambridge, UK), whereas those against human p21, cyclin D1, E-cadherin, vimentin, fibronectin, Janus N-terminal kinase (JNK)1/2, phosphorylated (p)-JNK1/2, p38, p-p38, phosphoinositide 3-kinase (PI3K), p-PI3K, AKT, p-AKT, Src, p-Src, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, used as the internal control) were purchased from Cell Signaling Technology (Danvers, MA, USA). Short interfering RNAs (siRNAs) that targeted TXNDC5 were obtained from GenePharma (Shanghai, China). Invitrogen (Carlsbad, CA, USA) was the supplier of TRIzol, Moloney murine leukemia virus reverse transcriptase (M-MLV-RT), and Lipofectamine 2000, whereas Roche (Penzberg, Germany) provided SYBR Green I Master kits. Unless otherwise mentioned, biochemical reagents used were from Sigma (St. Louis, MO, USA).
Wang H., Yang X., Guo Y., Shui L., Li S., Bai Y., Liu Y., Zeng M, & Xia J. (2019). HERG1 promotes esophageal squamous cell carcinoma growth and metastasis through TXNDC5 by activating the PI3K/AKT pathway. Journal of Experimental & Clinical Cancer Research : CR, 38, 324.
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10

Antibody Sourcing and Labeling

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Goat polyclonal antibodies against CL4 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, United States). Rabbit polyclonal antibodies were obtained from Abcam (Cambridge, MA, United States). The secondary antibodies were horseradish peroxidase (HRP)-conjugated anti-rabbit or anti-goat immunoglobulin (Ig) G (ZSGB-BIO, Beijing, China), Alexia Flour 488 (green)-labeled donkey anti-goat IgG (abcam, Cambridge, CA, United States).
Liang Z.Y., Kang X., Chen H., Wang M, & Guan W.X. (2017). Effect of Clostridium perfringens enterotoxin on gastric cancer cells SGC7901 which highly expressed claudin-4 protein. World Journal of Gastrointestinal Oncology, 9(4), 153-159.
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