The following primary and secondary antibodies were utilized: Gαi1/2 (anti-sera) [53 ], PDZ-RhoGEF (ab110059, abcam), HA (3724, Cell Signaling), FLAG (F1804, Sigma), P-MLC (3671, Cell Signaling), c-myc (13-2500, Invitrogen), Streptavidin-IRDye800 (925-32230, LI-COR), GFP (A11122, Invitrogen). Primary antibodies were made in 3% BSA and 0.1% Sodium azide and the blots were incubated in primary antibody overnight at 4 °C except 1 hr incubation at RT for Streptavidin-IRDye800. Secondary antibody goat anti-rabbit DyLight™ 800 (SA535571, Invitrogen), goat anti-mouse IRDye 800CW (926-32210, LICOR) at 1:10,000 dilution and goat anti-rabbit Alexa Fluor 488 (A11034, Invitrogen) at 1:1000 dilution.
Goat anti rabbit dylight 800
Manufactured by Thermo Fisher Scientific
Goat anti-rabbit DyLight 800 is a secondary antibody conjugated with the DyLight 800 fluorescent dye. It is designed to detect and visualize primary rabbit antibodies in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse dylight 680, by Thermo Fisher Scientific (6 mentions)
Goat anti mouse dylight 800, by Thermo Fisher Scientific (4 mentions)
C myc, by Thermo Fisher Scientific (3 mentions)
Streptavidin irdye800, by LI COR (3 mentions)
Irdye 800cw goat anti mouse, by LI COR (3 mentions)
Goat anti rabbit dylight680, by Thermo Fisher Scientific (3 mentions)
Gapdh antibody, by Merck Group (3 mentions)
Odyssey imaging system, by LI COR (3 mentions)
Nupage, by Thermo Fisher Scientific (3 mentions)
Flag tag (flag), by Thermo Fisher Scientific (2 mentions)
Ha c29f4, by Cell Signaling Technology (2 mentions)
Tubb6, by Proteintech (2 mentions)
Arhgap10, by Proteintech (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
Lysis buffer, by Merck Group (2 mentions)
Socs3, by Cell Signaling Technology (2 mentions)
P stat3, by Santa Cruz Biotechnology (2 mentions)
P mlc, by Cell Signaling Technology (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
13 protocols using goat anti rabbit dylight 800
1
Immunoblotting Protocol for Protein Detection
2
Multiparametric Immunofluorescence Analysis
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mouse anti-α-actinin (Sigma, Cat# A7811), rabbit anti-ANF (Millipore, Cat# AB-5490), mouse anti-GM130 (BD Transduction Laboratories, Cat# 610822), mouse M1 anti-Flag (Millipore Sigma, Cat# F3040) labelled with Alexa Fluor 488 using Thermofisher labeling kit A20181 (gift from Dr. Manojkumar Puthenveedu), rabbit anti-p-PKD (Cell Signaling Technology, Cat# 2051), rabbit anti-t-PKD (Cell Signaling Technology, Cat# 2054), rabbit anti-p-ERK (Cell Signaling Technology, Cat# 4370S), mouse anti-t-ERK (Cell Signaling Technology, Cat# 4696S), rabbit anti-p-HDAC (Cell Signaling Technology Cat# 3433), mouse anti-t-HDAC (Santa Cruz Biotechnology Cat# sc-133225), mouse anti-GAPDH (Invitrogen, Cat# MA5-15738), goat anti-rabbit DyLight 800 (Invitrogen, Cat# SA535571), goat anti-mouse IRDye 680RD (LICOR, Cat# 926-68070), goat anti-mouse 568 secondary antibody (Invitrogen, Cat# A-11031), goat anti-rabbit 568 secondary antibody (Invitrogen, Cat# A-11011), goat anti mouse 488 secondary antibody (Invitrogen, Cat# A-11029).
3
Immunoblotting with Fluorescent Antibodies
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Nitrocellulose membranes were blocked in blocking buffer: 5% milk (BLOT- QuickBlocker™ EMD Millipore WB57) in TBST followed by overnight incubation with primary antibody at 4 °C. Membranes were washed with TBST and incubated with secondary antibody for 1 h at room temperature, followed by washed with TBST. The membranes were imaged on Odyssey® CLx (LI-COR Biosciences) and quantification of bands was performed using Image Studio™ (LI-COR Biosciences).
Primary and secondary antibodies were prepared in blocking buffer.
Primary antibodies: Mouse anti-FLAG (1:1000; Sigma F1804); Rabbit anti-MATR3 (1:4000; Abcam ab151714); Mouse anti-V5 (1:1000; Invitrogen R960-25); Mouse anti- α tubulin (1:8000; Sigma T5168)
Secondary antibodies: Goat anti-mouse Dylight 680 (1:10000; LI-COR 925-68070); Goat anti-rabbit Dylight 680 (1:10000; Invitrogen 35568); Goat anti-mouse Dylight 800 (1:10000; Invitrogen SA5-10176); Goat anti-rabbit Dylight 800 (1:10000; Invitrogen SA5-35571)
Primary and secondary antibodies were prepared in blocking buffer.
Primary antibodies: Mouse anti-FLAG (1:1000; Sigma F1804); Rabbit anti-MATR3 (1:4000; Abcam ab151714); Mouse anti-V5 (1:1000; Invitrogen R960-25); Mouse anti- α tubulin (1:8000; Sigma T5168)
Secondary antibodies: Goat anti-mouse Dylight 680 (1:10000; LI-COR 925-68070); Goat anti-rabbit Dylight 680 (1:10000; Invitrogen 35568); Goat anti-mouse Dylight 800 (1:10000; Invitrogen SA5-10176); Goat anti-rabbit Dylight 800 (1:10000; Invitrogen SA5-35571)
4
Western Blot Antibody Immunodetection
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The following primary and secondary antibodies were used: Gαi1/2 (anti-sera) 58 (link), c-Myc (13–2500, Invitrogen), GFP (A11122, Invitrogen), HA (C29F4, Cell Signaling), FLAG (PA1–984B, Invitrogen). Streptavidin-IRDye800 was from LI-COR (925–32230). Primary antibodies were diluted in 3% bovine serum albumin (BSA) and 0.1% sodium azide and incubated with blots overnight at 4°C. Streptavidin-IRDye800 was incubated for 1 hour at room temperature. For secondary antibodies, goat anti-rabbit DyLight 800 (SA535571, Invitrogen) and goat anti-mouse IRDye 800CW (926–32210, LI-COR) were used at 1:10,000.
5
Quantitative Western Blotting Analysis
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The following primary and secondary antibodies were used: Gαi1/2 (anti-sera) (56 (link)), c-Myc (13-2500, Invitrogen), GFP (A11122, Invitrogen), HA (C29F4, Cell Signaling Technology), FLAG (PA1-984B, Invitrogen). Streptavidin-IRDye800 was from LI-COR (925-32230). Primary antibodies were diluted in 3% bovine serum albumin and 0.1% sodium azide and incubated with blots overnight at 4 °C. Streptavidin-IRDye800 was incubated for 1 h at room temperature (RT). For secondary antibodies, goat anti-rabbit DyLight 800 (SA535571, Invitrogen) and goat anti-mouse IRDye 800CW (926-32210, LI-COR) were used at 1:10,000.
6
Osteoclast Protein Expression Analysis
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Osteoclast lysates were prepared in lysis buffer (50 mM Tris pH7.4, 150 mM NaCl, 1% IGEPAL CA-630, 2 mM EDTA, 50 mM NaF, 10% glycerol, protease inhibitor cocktail). Culture supernatants were collected to assess the secretion of Cathepsin K. Protein concentrations were determined with Bradford. After resolution by SDS-PAGE, proteins were transferred to PVDF-FL membranes (Immobilon-FL Transfer Membrane) and analyzed by immunoblot. Primary antibodies were: TUBB6 (1:1000), ARHGAP10 (ProteinTech S5136AP, 1:1000), GAPDH (CellSignaling #2118, 1:5000), Sigma antibodies EB1 (E3406, 1:2500), K40-acetylated tubulin (T6793, 1:1000) and actin (A2103, 1:1000), Abcam antibodies Vinculin (ab108620, 1:5000), CLASP1 (ab108620, 1:5000) and Histone H3 (ab1791, 1:1000), total β tubulin (Developmental Studies Hybridoma Bank E7, 1:1000), Santa Cruz Biotechnologies antibodies TUBB5 (SAO.4G5, sc-58884, 1:5000) and Cathepsin K (E-7, sc-48353, 1:500). Secondary antibodies were: Goat-anti-mouse Dylight680 (Invitrogen, 35518), Goat-anti-mouse Dylight800 (Invitrogen, SA535521), Goat-anti-rabbit Dylight680 (Invitrogen, 35568), Goat-anti-rabbit Dylight800 (Invitrogen, SA535571). Signals were acquired using the Odyssey Infrared Imaging System (LI-COR Biosciences, United States).
7
Osteoclast Protein Immunoblot Analysis
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Osteoclast lysates were prepared in lysis buffer (50 mM Tris pH7.4, 150 mM NaCl, 1% IGEPAL CA-630, 2 mM EDTA, 50 mM NaF, 10% glycerol, protease inhibitor cocktail).
Protein concentrations were determined with Bradford. After resolution by SDS-PAGE, proteins were transferred to PVDF-FL membranes (Immobilon-FL Transfer Membrane) and analyzed by immunoblot. Primary antibodies were: TUBB6 (1:1000), ARHGAP10 (ProteinTech S5136AP, 1:1000), GAPDH (CellSignaling #2118, 1:5000), Sigma antibodies EB1 (E3406, 1:2500), K40-acetylated tubulin (T6793, 1:1000) and actin (A2103, 1:1000), Abcam antibodies Vinculin (ab108620, 1:5000), CLASP1 (ab108620, 1:5000) and Histone H3 (ab1791, 1:1000), total β tubulin (Developmental Studies Hybridoma Bank E7, 1:1000), preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. TUBB5 (SAO.4G5, Santa Cruz Biotechnologies sc-58884, 1:5000). Secondary antibodies were: Goat-anti-mouse Dylight680 (Invitrogen, 35518), Goat-anti-mouse Dylight800 (Invitrogen, SA535521), Goat-anti-rabbit Dylight680 (Invitrogen, 35568), Goat-anti-rabbit Dylight800 (Invitrogen, SA535571). Signals were acquired using the Odyssey Infrared Imaging System (LI-COR Biosciences, USA).
Protein concentrations were determined with Bradford. After resolution by SDS-PAGE, proteins were transferred to PVDF-FL membranes (Immobilon-FL Transfer Membrane) and analyzed by immunoblot. Primary antibodies were: TUBB6 (1:1000), ARHGAP10 (ProteinTech S5136AP, 1:1000), GAPDH (CellSignaling #2118, 1:5000), Sigma antibodies EB1 (E3406, 1:2500), K40-acetylated tubulin (T6793, 1:1000) and actin (A2103, 1:1000), Abcam antibodies Vinculin (ab108620, 1:5000), CLASP1 (ab108620, 1:5000) and Histone H3 (ab1791, 1:1000), total β tubulin (Developmental Studies Hybridoma Bank E7, 1:1000), preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. TUBB5 (SAO.4G5, Santa Cruz Biotechnologies sc-58884, 1:5000). Secondary antibodies were: Goat-anti-mouse Dylight680 (Invitrogen, 35518), Goat-anti-mouse Dylight800 (Invitrogen, SA535521), Goat-anti-rabbit Dylight680 (Invitrogen, 35568), Goat-anti-rabbit Dylight800 (Invitrogen, SA535571). Signals were acquired using the Odyssey Infrared Imaging System (LI-COR Biosciences, USA).
8
Western Blot Analysis of Noradrenergic Nuclei
9
Western Blot Analysis of Noradrenergic Nuclei
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Noradrenergic nuclei from the brainstem were homogenized in lysis buffer (Sigma Aldrich, St. Louis, MO) and protein concentrations were determined using BCA assay as previously described12 (link), 13 (link). 20μg of protein was loaded for each sample into SDS-PAGE gels (NuPAGE, Invitrogen, Carlsbad, CA). Proteins were then transferred onto PVDF membranes which were probed with antibodies including pSTAT-3 (1:1000; goat-polyclonal; Santa Cruz Biotechnology, Dallas, TX), SOCS-3 (1:1000; goat-polyclonal; Cell Signaling Technology, Danvers, MA), and GAPDH antibody (1:2000; mouse-monoclonal; Sigma-Aldrich, St. Louis, MO). After washing 3 times of 10 min each, the membranes were incubated in blocking solution containing goat anti-rabbit DyLight 800 and goat anti-mouse DyLight 680 secondary antibodies (1:5000; Thermo Fisher Scientific; Waltham, MA). Bands were visualized and analyzed using an Odyssey imaging system (Li-COR biosciences, Lincoln, NE).
10
Comprehensive Antibody Profiling for Cellular Analysis
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The antibodies used in this study were as follows: TGN46 (Abcam; ab16052); CCM3 (Acris; AP26023PU-N); BrdU (BD Biosciences; 555627), Lamp-2 (BD Biosciences; 555803), p21CIP1 (F-5) (Santa Cruz; sc-6246), p16INK4 (BD Biosciences; 511325), p62 (BD Biosciences; 610832); GAPDH (Calbiochem; CB-1001); LC3B (Cell Signaling; 3868), mTOR (Cell Signaling; 2983); H2AX (Ser139) (Millipore; 05-636); IL-8 (500-P28) (PeproTech; 500-P28); NFκB p65 (Santa Cruz; sc-8008); p53 (DO-1) (Santa Cruz; sc-126); C/EBPβ (Santa Cruz; sc-150); and tubulin (T5168) (Sigma-Aldrich). The secondary antibodies used were as follows: goat anti-rabbit DyLight™ 800, goat anti-mouse DyLight™ 680 (Thermo Scientific); goat anti-mouse Alexa 488, goat anti-rabbit Alexa 488, goat anti-mouse Alexa 594, and goat anti-rabbit Alexa 546 (Molecular Probes).
All plasmids were constructed using standard molecular biology techniques.
All plasmids were constructed using standard molecular biology techniques.
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