ELISA plates were coated with 5 ng/well of OVA or PE conjugates or unconjugated protein as control in carbonate buffer at pH 9.6 and blocked with 1% gelatin. Primary antibodies were incubated with anti-mouse IgG antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA) to measure hapten-specific serum IgG antibody titers as described previously [17 (link)]. We have previously shown that vaccination elicits hapten-specific serum antibodies that cross-react with the target free drug [17 (link),23 ,24 (link)].
Anti mouse igg antibody
Manufactured by Jackson ImmunoResearch
Sourced in United States, United Kingdom
Anti-mouse IgG antibody is a laboratory reagent used to detect and quantify mouse immunoglobulin G (IgG) in various experimental and analytical applications. It is a secondary antibody that binds specifically to mouse IgG, allowing for the identification and measurement of mouse IgG in samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti hsc70 antibody mab2191, by Abnova (2 mentions)
C digit blot scanner and image studio for c digit, by LI COR (2 mentions)
Anti rat igg, by Thermo Fisher Scientific (2 mentions)
Alkaline phosphatase conjugated anti human igg antibody, by Jackson ImmunoResearch (2 mentions)
Peroxidase conjugated goat anti rabbit, by Jackson ImmunoResearch (1 mentions)
Mannan, by Merck Group (1 mentions)
Low viscosity carboxy methyl cellulose, by Merck Group (1 mentions)
Fatty acid free bovine serum albumin, by Merck Group (1 mentions)
α tubulin monoclonal antibody, by Cell Signaling Technology (1 mentions)
Nupage 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions)
C digit blot scanner, by LI COR (1 mentions)
Image studio for c digit, by LI COR (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Merck Group (1 mentions)
Peroxidase goat anti human igg antibody, by Jackson ImmunoResearch (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Bradford reagent, by Merck Group (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Abc elite dab peroxidase detection system, by Vector Laboratories (1 mentions)
Anti ulbp1, by R&D Systems (1 mentions)
18 protocols using anti mouse igg antibody
1
ELISA for Hapten-Specific Antibody Titers
2
Affinity Purification of Viral Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hemagglutinin (HA) peptide, anti-HA agarose, and rabbit HA epitope tag antibody were purchased from Pierce, Waltham, Massachusetts, USA. Peroxidase-conjugated goat anti-rabbit and anti-mouse IgG antibodies were purchased from Jackson ImmunoResearch, Cambridge, UK. Both mouse monoclonal anti-nucleoprotein (NP) (clone ZDD4) and anti-VP40 (clone 9B2-F2) were produced in-house and diluted at 1:500 and 1:100, respectively. Low viscosity carboxymethylcellulose, mannan, and fatty acid-free bovine serum albumin (BSA) were purchased from Sigma Aldrich, St Louis, MO, USA. True Blue peroxidase substrate was purchased from Seracare KPL (Milford, MA, USA). Protein low binding (LoBind) 1.5 mL tubes were purchased from Eppendorf France SAS (Montesson, France) and 1 M N-2-Hydroxyethylpiperazine-N-2-Ethane Sulfonic Acid (HEPES) solution was from Invitrogen (Carlsbad, CA, USA).
3
PBMC Lysate SDS-PAGE and Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMC lysates were subjected to SDS-PAGE using NuPAGE 4–12% Bis-Tris Gels (Invitrogen). The procedure was described previously [20 (link)]. Primary antibodies used are Orai1 polyclonal antibody (ProSci #4041) and α-Tubulin monoclonal antibody (Cell Signaling Technology #2144). Both HRP-conjugated goat anti-rabbit and anti-mouse IgG antibodies were obtained from Jackson ImmunoResearch Laboratories.
4
Western Blotting Analysis of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting analysis was conducted as previously reported [19 (link)]. The primary antibodies used were anti-HSC70 antibody (MAB2191; Abnova, Taipei, Taiwan), anti-CD63 antibody (sc-5725; Santa Cruz Biotechnology, Texas, USA), and anti-CYCS antibody (sc-13156), and the secondary antibodies used were horseradish peroxidase conjugated anti-rat IgG antibodies (81–9520; Invitrogen) and anti-mouse IgG antibodies (115-035-044; Jackson ImmunoResearch, West Grove, PA, USA). Signals were visualized using an Immunostar LD Kit (Wako, Tokyo, Japan) and the C-DiGit Blot Scanner and Image Studio for C-DiGit (LI-COR, Lincoln, NE, USA) according to the manufacturer’s protocols.
5
Western Blot Analysis of Exosomal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were isolated from the exosomal fraction and MGCs using TRIZOL Reagent (Life Technologies) according to the manufacturer′s protocol, and were
resuspended in 8 M urea buffer (8 M urea, 10 mM dithiothreitol, 50 mM Tris(hydroxymethyl)aminomethane, pH 8.0). Laemmli buffer [26 (link)] was added to each sample and immediately boiled for 5 min. Western blot analysis was conducted as previously reported [27 (link)]. The primary antibodies used were anti-HSC70 antibody (MAB2191; Abnova, Taipei, Taiwan), anti-ACTB antibody (GTX109639; GeneTex, CA,
USA), and anti-CYCS antibody (sc-13156; Santa Cruz Biotechnology, Texas, USA), and the secondary antibodies used were horseradish peroxidase conjugated
anti-rabbit IgG (AP132P; Merck Millipore), anti-rat IgG (81-9520; Invitrogen), and anti-mouse IgG antibodies (115-035-044; Jackson ImmunoResearch, West Grove,
PA, USA). Signals were visualized using an Immunostar LD Kit (Wako, Tokyo, Japan) and the C-DiGit Blot Scanner and Image Studio for C-DiGit (LI-COR, Lincoln,
NE, USA) according to the manufacturer′s protocols.
resuspended in 8 M urea buffer (8 M urea, 10 mM dithiothreitol, 50 mM Tris(hydroxymethyl)aminomethane, pH 8.0). Laemmli buffer [26 (link)] was added to each sample and immediately boiled for 5 min. Western blot analysis was conducted as previously reported [27 (link)]. The primary antibodies used were anti-HSC70 antibody (MAB2191; Abnova, Taipei, Taiwan), anti-ACTB antibody (GTX109639; GeneTex, CA,
USA), and anti-CYCS antibody (sc-13156; Santa Cruz Biotechnology, Texas, USA), and the secondary antibodies used were horseradish peroxidase conjugated
anti-rabbit IgG (AP132P; Merck Millipore), anti-rat IgG (81-9520; Invitrogen), and anti-mouse IgG antibodies (115-035-044; Jackson ImmunoResearch, West Grove,
PA, USA). Signals were visualized using an Immunostar LD Kit (Wako, Tokyo, Japan) and the C-DiGit Blot Scanner and Image Studio for C-DiGit (LI-COR, Lincoln,
NE, USA) according to the manufacturer′s protocols.
6
Western Blot Analysis of EV Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The EV‐enriched fraction and MGCs were resuspended in Laemmli buffer (Laemmli, 1970 ) and immediately boiled for 5 min. Western blot analysis was conducted as reported previously (Matsuno et al., 2019 ). The primary antibodies used were anti‐HSC70 rat antibody (MAB2191; Abnova), anti‐ALIX rabbit antibody (ab186429; Abcam), and anti‐CYCS mouse antibody (sc‐13156; Santa Cruz Biotechnology) as well‐known EV markers. The secondary antibodies used were a horseradish peroxidase‐conjugated anti‐rabbit IgG (AP132P; Merck), anti‐rat IgG (81‐9520; Thermo Fisher Scientific), and anti‐mouse IgG antibodies (115‐035‐044; Jackson ImmunoResearch). Signals were visualized using an Immunostar LD Kit (FUJIFILM Wako Pure Chemical Corporation) and the C‐DiGit Blot Scanner and Image Studio for C‐DiGit (LI‐COR).
7
Western Blot Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were collected and resuspended in Laemmeli buffer. Proteins were separated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membranes. After incubation with primary antibody the membrane was incubated with and goat horseradish peroxidase-conjugated anti-rabbit IgG (Sigma A0545, 1/10 000) or anti-mouse IgG antibodies (Jackson ImmunoResearch 61871, 1/10 000).
8
Genotyping and Platelet Antigen Detection Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For genotyping, genome DNA was extracted from cells using a QuickGene DNA whole blood kit (DB-S, Kurabo Biomedical). The extracted DNA was amplified using multiplex polymerase chain reaction (PCR), hybridized to oligonucleotide probes immobilized on microsphere beads, labeled with SAPE using WAKFlow HPA typing reagents (4R408, Wakunaga Pharmaceutical), and analyzed using Luminex XYP.
For the modified rapid monoclonal antibody–specific immobilization of platelet antigens (MR-MAIPA) assay,16 (link) platelet samples were added to anti–HPA-1a sera or negative control serum and capture antibodies: anti-hCD41 (clone HIP8; Biolegend, #303702), anti-hCD41 (clone P2; Beckman Coulter, PN IM0145), or anti-hCD61 (clone SZ21; Beckman Coulter, IM0540). After lysing the platelets, samples were applied to anti-mouse IgG antibody–coated plates, washed, mixed with peroxidase–goat-anti-human IgG antibody (Jackson ImmunoResearch Laboratories, #109-035-088), and subjected to fluorescence by the addition of a substrate (KPL, #50-76-00). The reaction was then stopped by the addition of 3, 3′, 5, 5′-tetramethylbenzidine stop solution (KPL, #50-85-05), and the absorbance was measured at 450 nm (MTP-120, Hitachi, Tokyo, Japan).
For the modified rapid monoclonal antibody–specific immobilization of platelet antigens (MR-MAIPA) assay,16 (link) platelet samples were added to anti–HPA-1a sera or negative control serum and capture antibodies: anti-hCD41 (clone HIP8; Biolegend, #303702), anti-hCD41 (clone P2; Beckman Coulter, PN IM0145), or anti-hCD61 (clone SZ21; Beckman Coulter, IM0540). After lysing the platelets, samples were applied to anti-mouse IgG antibody–coated plates, washed, mixed with peroxidase–goat-anti-human IgG antibody (Jackson ImmunoResearch Laboratories, #109-035-088), and subjected to fluorescence by the addition of a substrate (KPL, #50-76-00). The reaction was then stopped by the addition of 3, 3′, 5, 5′-tetramethylbenzidine stop solution (KPL, #50-85-05), and the absorbance was measured at 450 nm (MTP-120, Hitachi, Tokyo, Japan).
9
Quantitative Western Blot Analysis of Transglutaminase
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HEK293 cells expressing hTG2, zTGs2, or the non-transfected negative control cells were lysed in lysis buffer (Tris-HCl 20 mM pH 8, NaCl 500 mM, 1% Triton X-100, supplemented with cOmplete™ Protease Inhibitor Cocktail Tablets), sonicated, and centrifuged at 14,000× g for 15 min at 4 °C. The clear protein lysates were collected, quantified with Bradford reagent (Sigma-Aldrich, cat#B6916), the proteins were resolved on SDS-polyacrylamide gel and transferred onto a nitrocellulose membrane. After a blocking step of 45 min with 5% skim milk in PBS at room temperature (RT), the membrane was incubated for 1 h with an anti-SV5 antibody. After three washes with PBS-Tween 20 0.1% and three with PBS, the membrane was incubated with anti-mouse IgG antibody (Jackson ImmunoResearch, Ely, UK) diluted in the blocking buffer in volumetric ratio 1:2500. After three washes with PBS-Tween 20 0.1% and three with PBS, the membrane was developed with Pierce™ ECL Western Blotting Substrate (Thermo Scientific, cat#32209) and imaged with ChemiDoc™ MP Imaging System (Bio-Rad).
10
Serological Assay for Pandemrix Antigens
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polystyrene plates were coated with rabbit antibodies to HA, NA (anti-HA MAb (11055-RM05), anti-NA MAb (11058-R001) and anti-NP PAb (11675-RP01) all from Sino Biological, China), 1 µg/mL in PBS (100 µl/well), and kept overnight at +4°C 0.5% BSA-PBS was used for post-coating. After washing with PBS, Pandemrix antigen suspension (100 µl/well) was added and incubated in the antibody coated wells. After washing, mouse monoclonal antibodies to HA, NA or NP (anti-HA MAb (11055-MM08), anti-NA Mab (11058-MM07) or anti- NP MAb (11675-MM03) (1 µg/ml in 0.5% BSA-PBS) were added. In the EIA for IgG-antibodies to Pandemrix derived NP, plasma samples were diluted 1∶300 in 0.5% bovine serum albumin (BSA)-PBS. The binding to antibodies was detected with alkaline phosphatase conjugated anti-human IgG-antibody or anti-mouse IgG-antibody (both from Jackson ImmunoResearch, USA). After adding the substrate the color reaction was read.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!