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Ab40873

Manufactured by Abcam
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Sourced in United Kingdom, United States, Switzerland

Ab40873 is a recombinant monoclonal antibody produced in mice. It is designed to detect the presence of the target protein in biological samples.

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Lab products found in correlation

Ab106926, by Abcam (2 mentions) Ab181153, by Abcam (2 mentions) Ab90716, by Abcam (2 mentions) Alexa fluor 488 igg 53 5381 80, by Thermo Fisher Scientific (2 mentions) Igg h l rhodamine red conjugate r 6394, by Thermo Fisher Scientific (2 mentions) Hoechst, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 conjugate, by Thermo Fisher Scientific (2 mentions) Ab40879, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) H l alexa fluor 488, by Abcam (1 mentions) Dmi8 inverted tiling microscope, by Leica (1 mentions) Secondary antibodies conjugated to horseradish peroxidase, by Santa Cruz Biotechnology (1 mentions) Anti actin 1 19, by Santa Cruz Biotechnology (1 mentions) Sc 21701, by Santa Cruz Biotechnology (1 mentions) Mouse anti muc5ac, by Santa Cruz Biotechnology (1 mentions) Goat anti notch1, by Santa Cruz Biotechnology (1 mentions) Sc 1616, by Santa Cruz Biotechnology (1 mentions) Ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Ab5450, by Abcam (1 mentions)

6 protocols using ab40873

1

Immunofluorescence Staining of Lung Tissue

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Formalin-fixed, paraffin-embedded tissue sections were deparaffinized in xylene, rehydrated, and stained with antiuteroglobin antibody for club cells (1:1,600) (Abcam, ab40873) or antiprosurfactant protein C antibody for alveoli type II (1:1,000) (Abcam, ab40879). Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (Abcam, ab150077) was used as secondary antibody (1:1,000). The nucleus was stained with DAPI (Molecular Probes). Images of three sections per mouse, 100 µm apart, were acquired with Leica DMi8 inverted tiling microscope (Leica Microsystems) and processed using LAS X.
Hariharan V.N., Shin M., Chang C.W., O’Reilly D., Biscans A., Yamada K., Guo Z., Somasundaran M., Tang Q., Monopoli K., Krishnamurthy P.M., Devi G., McHugh N., Cooper D.A., Echeverria D., Cruz J., Chan I.L., Liu P., Lim S.Y., McConnell J., Singh S.P., Hildebrand S., Sousa J., Davis S.M., Kennedy Z., Ferguson C., Godinho B.M., Thillier Y., Caiazzi J., Ly S., Muhuri M., Kelly K., Humphries F., Cousineau A., Parsi K.M., Li Q., Wang Y., Maehr R., Gao G., Korkin D., McDougall W.M., Finberg R.W., Fitzgerald K.A., Wang J.P., Watts J.K, & Khvorova A. (2023). Divalent siRNAs are bioavailable in the lung and efficiently block SARS-CoV-2 infection. Proceedings of the National Academy of Sciences of the United States of America, 120(11), e2219523120.
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2

Lung Tissue Protein Expression Analysis

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Lung tissue (200 mg) was homogenized in RIPA lysis buffer (100 ml PBS 1x pH 7.2, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, supplemented with protease inhibitors). Total protein was quantified using Bradford assay (Bio-Rad, Hercules, CA). Samples were subjected to SDS-PAGE electrophoresis, using a 15% acrylamide gel for CC10 and 10% acrylamide for Notch1. Seventy five μg of protein were loaded for CC10 and 50 μg for Notch1. They were transferred to PVDF membranes and blocked with 5% nonfat milk. The following primary antibodies were used: mouse anti-Muc5ac (1:2000, sc-21701, Santa Cruz Biotechnology), goat anti-Muc5b (1:2000, sc-135508, Santa Cruz Biotechnology), rabbit anti-CC10 (1:4000, ab40873, Abcam, UK), goat anti-Notch1 (1:2000, sc-6015, Santa Cruz Biotechnology), and Anti-Actin I-19 (1:2000, sc-1616, Santa Cruz Biotechnology, USA). Secondary antibodies conjugated to horseradish peroxidase (Santa Cruz Biotechnology) were used. Brain and stomach were used as positive controls. Water without antibodies was used as negative control. Proteins were detected by Pierce, ECL Western Blotting Substrate (Pierce Biotechnology, Rockford, IL). Images were film-captured. Density of band was measured using Fiji software (ImageJ 2.0.0-rc-54/1.51h) and protein amounts were normalized with the actin signal.
Méndez A., Rojas D.A., Ponce C.A., Bustamante R., Beltrán C.J., Toledo J., García-Angulo V.A., Henriquez M, & Vargas S.L. (2019). Primary infection by Pneumocystis induces Notch-independent Clara cell mucin production in rat distal airways. PLoS ONE, 14(6), e0217684.
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3

Immunofluorescence Staining of Lung Cells

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Briefly, tissues were stained with goat polyclonal IgG to Irp2 (ab106926, Abcam) at a 1:50 dilution or rabbit monoclonal IgG to Irp2 (ab181153, Abcam) at a 1:50 dilution. Co-staining for type II epithelial cells was carried out using a rabbit polyclonal IgG to prosurfactant protein C (Pro-SPC) (ab90716, Abcam) at a 1:50 dilution and for type I epithelial cells using anti-mouse podoplanin alexa Fluor® 488 IgG (53-5381-80, Affymetrix) at a 1:500 dilution. Co-staining for Irp2 and markers of ciliated airway epithelial cells was conducted using a rabbit IgG for acetyl-α-tubulin (Lys40) (D20G3, Cell Signaling) at a 1:50 dilution, and co staining for non-ciliated secretory epithelial cells was conducted using a rabbit polyclonal IgG to uteroglobin (ab40873, Abcam) at a 1:100 dilution. Secondary staining was carried out using goat anti-rabbit IgG (H+L) rhodamine red conjugate (R-6394, Life Technologies) at a 1:500 dilution or donkey anti-goat IgG (H+L) secondary antibody, alexa Fluor® 488 conjugate (A-11055, Life technologies) at a 1:500 dilution. Nuclei were counter-stained using TO-PRO®-3 Iodide (T3605, Life Technologies) at a 1:1000 dilution or Hoechst (33342, ThermoFisher Scientific) at a 1:300 dilution.
Cloonan S.M., Glass K., Laucho-Contreras M.E., Bhashyam A.R., Cervo M., Pabón M.A., Konrad C., Polverino F., Siempos I.I., Perez E., Mizumura K., Ghosh M.C., Parameswaran H., Williams N.C., Rooney K.T., Chen Z.H., Goldklang M.P., Yuan G.C., Moore S.C., Demeo D.L., Rouault T.A., D’Armiento J.M., Schon E.A., Manfredi G., Quackenbush J., Mahmood A., Silverman E.K., Owen C.A, & Choi A.M. (2016). Mitochondrial iron chelation ameliorates cigarette-smoke induced bronchitis and emphysema in mice. Nature medicine, 22(2), 163-174.
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4

Immunofluorescence Staining of Lung Organoids

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The organoids were then formalin fixed and paraffin embedded. Sections were deparaffinized with xylene, treated with antigen retrieval solution (1 g NaOH, 2.1 g citric acid in 1 L of H2O) for 20 min in steam, cooled to room temperature, permeabilized with PBS-0.2% Triton for 15 min, and blocked with 10% donkey serum in PBS. Sections were stained with monoclonal rabbit antithyroid transcription factor 1 (TTF1, ab76013; Abcam, Cambridge, MA) or rabbit anti-CC10 (ab40873, Abcam, Cambridge, MA, USA) at 1:500 followed by Alexa 594-conjugated donkey antirabbit secondary antibody (ab150076, Abcam, Cambridge, MA, USA) at 1:500, goat anti-GFP (ab5450; Abcam, Cambridge, MA, USA) at 1:500 followed by Alexa 488-conjugated donkey anti-goat secondary antibody (ab150129, Abcam, Cambridge, MA, USA) at 1:500. Images were taken at 10×, 20×, and 40× with an Olympus FV1000 confocal microscope. Image analysis were performed using ImageJ software.
Ciechanowicz A.K., Sielatycka K., Cymer M., Skoda M., Suszyńska M., Bujko K., Ratajczak M.Z., Krause D.S, & Kucia M. (2021). Bone Marrow-Derived VSELs Engraft as Lung Epithelial Progenitor Cells after Bleomycin-Induced Lung Injury. Cells, 10(7), 1570.
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5

Immunofluorescence Staining of Lung Cells

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Briefly, tissues were stained with goat polyclonal IgG to Irp2 (ab106926, Abcam) at a 1:50 dilution or rabbit monoclonal IgG to Irp2 (ab181153, Abcam) at a 1:50 dilution. Co-staining for type II epithelial cells was carried out using a rabbit polyclonal IgG to prosurfactant protein C (Pro-SPC) (ab90716, Abcam) at a 1:50 dilution and for type I epithelial cells using anti-mouse podoplanin alexa Fluor® 488 IgG (53-5381-80, Affymetrix) at a 1:500 dilution. Co-staining for Irp2 and markers of ciliated airway epithelial cells was conducted using a rabbit IgG for acetyl-α-tubulin (Lys40) (D20G3, Cell Signaling) at a 1:50 dilution, and co staining for non-ciliated secretory epithelial cells was conducted using a rabbit polyclonal IgG to uteroglobin (ab40873, Abcam) at a 1:100 dilution. Secondary staining was carried out using goat anti-rabbit IgG (H+L) rhodamine red conjugate (R-6394, Life Technologies) at a 1:500 dilution or donkey anti-goat IgG (H+L) secondary antibody, alexa Fluor® 488 conjugate (A-11055, Life technologies) at a 1:500 dilution. Nuclei were counter-stained using TO-PRO®-3 Iodide (T3605, Life Technologies) at a 1:1000 dilution or Hoechst (33342, ThermoFisher Scientific) at a 1:300 dilution.
Cloonan S.M., Glass K., Laucho-Contreras M.E., Bhashyam A.R., Cervo M., Pabón M.A., Konrad C., Polverino F., Siempos I.I., Perez E., Mizumura K., Ghosh M.C., Parameswaran H., Williams N.C., Rooney K.T., Chen Z.H., Goldklang M.P., Yuan G.C., Moore S.C., Demeo D.L., Rouault T.A., D’Armiento J.M., Schon E.A., Manfredi G., Quackenbush J., Mahmood A., Silverman E.K., Owen C.A, & Choi A.M. (2016). Mitochondrial iron chelation ameliorates cigarette-smoke induced bronchitis and emphysema in mice. Nature medicine, 22(2), 163-174.
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6

Antibody Immunodetection Protocol

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The following antibodies were used: rabbit anti-human transferrin (A0061, Dako); rabbit anti-human Gc-globulin (also known as VDBP, A0021, Dako); rabbit anti-uteroglobin (also known as CC16, ab40873, Abcam); rabbit anti-SLC1A5 (also known as SGLT2, ab84903, Abcam); rabbit NaPi-IIa (gift from C.A.Wagner, University of Zurich, Zurich, Switzerland); rabbit anti-human AQP1 (ab2219, Millipore); mouse anti-β-actin (A2228, Sigma-Aldrich); mouse conjugated to Fluorescein (FITC) anti-PI(4,5)P2 (Z-G045, Echelon Biosciences Inc.); mouse anti-EEA1 (610456, BD Bioscience); rabbit anti-RFP (600–401-379, ROCKLAND); sheep anti-LRP2 (gift from P. Verroust and R. Kozyraki, INSERM, Paris, France); mouse anti Flotillin-1(610821, BD Bioscience); mouse anti-α-tubulin (T5168, Sigma-Aldrich); rabbit anti-GAPDH (2118, Cell Signaling Technology); rat anti-LAMP1 (sc-19992, Santa Cruz Biotechnology); goat anti-Cathepsin-D (Cts-D; sc-6486, Santa Cruz Biotechnology); rabbit anti-EGFR (1005 sc-03, Santa Cruz Biotechnology); Alexa-488 Phalloidin (F-actin, A12379, Thermofisher Scientific); mouse anti-Transferrin Receptor Antibody (H68,4, ThermoFisher Scientific); WGA FITC Conjugate (L 4895, Sigma-Aldrich); mouse anti-Na+/K+-ATPase subunit α1 (C464.6 EMD Millipore); rabbit anti-MPR and rabbit anti-OCRL (gift from A. De Matteis, Telethon Institute of Genetics and Medicine (TIGEM), Pozzuoli, Italy).
Festa B.P., Berquez M., Gassama A., Amrein I., Ismail H.M., Samardzija M., Staiano L., Luciani A., Grimm C., Nussbaum R.L., De Matteis M.A., Dorchies O.M., Scapozza L., Wolfer D.P, & Devuyst O. (2018). OCRL deficiency impairs endolysosomal function in a humanized mouse model for Lowe syndrome and Dent disease. Human Molecular Genetics, 28(12), 1931-1946.
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