We utilized the following reagents to investigate the effects of chronic, low-dose-rate 137Cs-γ radiation on NGF-induced neurite extension in PC12 cells: NGF 2.5S (N-100, Alomone Labs, Jerusalem, Israel); anti-ERK1/ERK2 mouse-monoclonal antibody (MAB1576) and anti-phosphorylated ERK1/ERK2 rabbit-polyclonal antibody (MAB1018, both R & D systems, Minneapolis, MN, USA); anti-Trk A rabbit polyclonal antibody (sc-118) and anti-phosphorylated Trk A mouse-monoclonal antibody (sc-8058, both Santa Cruz Biotechnology, Dallas, TX, USA)—Trk A is a high-affinity nerve growth factor receptor; anti-tyrosine hydroxylase rabbit-monoclonal antibody (ab137869); anti-Akt rabbit polyclonal antibody (ab8805) and anti-phosphorylated Akt rabbit monoclonal antibody (ab81283); anti-Ca2+/calmodulin-dependent kinase II (CaMKII) rabbit polyclonal antibody (ab131468) and anti-phosphorylated CaMKII rabbit polyclonal antibody (ab5683, all Abcam plc, Cambridge, UK); KN-62 (I2142, Sigma-Aldrich, St Louis, MO, USA).
Mab1018
Manufactured by R&D Systems
Sourced in United States
The MAB1018 is a mouse monoclonal antibody product from R&D Systems. It recognizes and binds to an undisclosed target antigen.
Automatically generated - may contain errors
Lab products found in correlation
Ab137869, by Abcam (1 mentions)
Ab5683, by Abcam (1 mentions)
Ab8805, by Abcam (1 mentions)
Kn 62, by Merck Group (1 mentions)
Ab81283, by Abcam (1 mentions)
Ab194806, by Abcam (1 mentions)
Ab39163, by Abcam (1 mentions)
Mab293, by R&D Systems (1 mentions)
Sc 7148, by Santa Cruz Biotechnology (1 mentions)
Sc 136548, by Santa Cruz Biotechnology (1 mentions)
Sc 13160, by Santa Cruz Biotechnology (1 mentions)
Sc 8439, by Santa Cruz Biotechnology (1 mentions)
Facscalibur, by BD (1 mentions)
Mab200, by R&D Systems (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
H 1200, by Vector Laboratories (1 mentions)
4 protocols using mab1018
1
Chronic Low-Dose Radiation Effects on PC12 Cells
2
Antibody Characterization for Western Blot
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The antibodies used for Western blot analysis in the present study were as follows: rabbit monoclonal anti-phospho-extracellular signal-regulated kinase (ERK)1/2 antibody (1:1,500, MAB1018, R&D Systems, Minneapolis, MA, United States), mouse monoclonal anti-phospho-p65 subunit of nuclear factor-kappa B (NF-κB) antibody (1:500, sc-136548, Santa Cruz Biotechnology Inc., Santa Cruz, CA, United States), rabbit polyclonal anti-phospho vascular endothelial growth factor receptor (VEGFR-2) antibody (1:1,000, ab194806, Abcam, Cambridge, United Kingdom), mouse monoclonal anti-intercellular adhesion molecule-1 (ICAM-1) antibody (1:1,000, sc-8439, Santa Cruz Biotechnology Inc.), mouse monoclonal anti-vascular cell adhesion protein-1 (VCAM-1) antibody (1:1,000, sc-13160, Santa Cruz Biotechnology Inc.), mouse monoclonal anti-VEGF antibody (1:1,500, MAB293, R&D Systems), rabbit polyclonal caspase-3 antibody (1:1,000, sc-7148, Santa Cruz Biotechnology Inc.), rabbit polyclonal ADAM-17 antibody (1:1,000, ab39163, Abcam), and rabbit monoclonal anti-TIMP-3 antibody (1:1,000, ab277794, Abcam). The recombinant proteins used in the present study were as follows: human TIMP-3 (Cat No 973-TM-010, R&D Systems), human tumor necrosis factor-alpha (TNF-α) (Cat No 210-TA, R&D Systems), and human VEGF (Cat No 293-VE-050, R&D Systems).
3
Quantifying Activated and Pan β1-Integrin on Cell Surface
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To determine the level of β1-integrin (activated or pan) at the plasma membrane, cells were trypsinized and washed two times in cold FACS buffer (PBS 2% serum), incubated with antibodies against activated (9EG7, 1/50) or pan (DF7 or MB1.2, 1/100) β1-integrin in cold FACS buffer. After incubation on ice, cells were washed and incubated on ice with PE or APC conjugates. When phospho-ERK1/2 was being analysed, 1 × 106 cells were fixed in 4% PFA for 10 min at 37 °C and then permeabilized in 90% ice-cold methanol for 30 min on ice. Cells were incubated with a phospho-ERK1/2 antibody (R&D Systems (MAB1018)) at 20 μg ml−1 for 1 h. After incubation on ice, cells were washed and incubated on ice with a BD Phosflow PE anti-rabbit secondary antibody (1:5). Flow cytometry data were acquired on a FACS Calibur (Becton Dickinson).
4
Docetaxel-Induced ERK1/2 and IL-1A Responses
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SUM-159 cell lines were cultured overnight in eight chambered cell culture slides (Corning 354118) pre-coated with Poly-L-lysine (Sigma P1399). Cells were then treated in the presence or absence of docetaxel (4 ng/ml) for 48 h and then stained for immunofluorescence. In brief, cells were fixed with 4% paraformaldehyde and permeabilized with 100% methanol. After blocking, cells were stained for Phospho ERK1/2 (R&D MAB1018) overnight at 4 C. The next day, cells were washed and stained with goat anti-rabbit Alexa Fluor 546 (ThermoFisher A-11035). Afterwards, cells were washed and stained for IL-1A (R&D MAB200) followed by goat anti-mouse Alexa Fluor 488 (ThermoFisher A-11001). Slides were mounted with Vectashield (Vector Labs H-1200 from Burlingame, Ca) and sealed with coverslips. Slides were imaged on the EVOS FL Auto Imaging System.
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