Log In Sign up
The largest database of trusted experimental protocols

Mouse igg2b

Manufactured by Merck Group
Visit Supplier
Sourced in United States, Macao

Mouse IgG2b is a type of immunoglobulin (antibody) produced by mouse B cells. It functions as a tool for research and scientific applications.

Automatically generated - may contain errors

Lab products found in correlation

Lsm 700, by Zeiss (2 mentions) Biotinylated horse anti mouse igg, by Vector Laboratories (1 mentions) Mouse igg2a, by BioLegend (1 mentions) Recombinant human m csf, by R&D Systems (1 mentions) Anti ifnar antibody, by PBL Assay Science (1 mentions) Ifn γ, by BD (1 mentions) Axiophoto, by Zeiss (1 mentions) Alexa 647 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Mouse igg1, by Developmental Studies Hybridoma Bank (1 mentions) Alexa fluor 555 conjugated α bungarotoxin, by Thermo Fisher Scientific (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Hoechst 33258, by Merck Group (1 mentions) Mab3656, by R&D Systems (1 mentions) Pe conjugated mouse igg2b, by R&D Systems (1 mentions) Rdnase, by Thermo Fisher Scientific (1 mentions) Metamorph software, by Molecular Devices (1 mentions) Celltiter 96 aqueous assay kit, by Promega (1 mentions) Low tox m rabbit complement, by Cedarlane (1 mentions)

6 protocols using mouse igg2b

1

Macrophage Differentiation and Stimulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human recombinant M-CSF (R&D Systems) was used for differentiation of blood monocytes into monocyte-derived macrophages (MDMs). Human recombinant IFN-β (BD Biosciences) and IFN-γ (BD Biosciences) were used for macrophage stimulations at the concentrations indicated. Anti-IFNAR antibody (PBL Assay Science) and corresponding isotype antibody mouse IgG2a (BioLegend) were used for neutralization experiments. Immunoperoxidase staining was performed with NUPR1 antibody (Abbexa), corresponding isotype antibody mouse IgG2b (Sigma), CD3 antibody (BD Pharmingen) and Biotinylated horse anti-mouse IgG (Vector).
R. Andrade P., Mehta M., Lu J., M. B. Teles R., Montoya D., O. Scumpia P., Nunes Sarno E., Ochoa M.T., Ma F., Pellegrini M, & Modlin R.L. (2019). The cell fate regulator NUPR1 is induced by Mycobacterium leprae via type I interferon in human leprosy. PLoS Neglected Tropical Diseases, 13(7), e0007589.
+ Open protocol
+ Expand
2

Immunofluorescence Staining of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were fixed in 4 % paraformaldehyde for 15–25 min at room temperature. After blocking in blocking buffer (PBS containing 5 FBS and 0.3 % Triton X-100), the cells were incubated with primary antibodies overnight at 4 °C. The cells were then rinsed with PBS three times and incubated with Alexa 488-, Alexa 555-, or Alexa 647-conjugated secondary antibodies (Thermo) for 1 h at room temperature. Nuclei were stained with 10 μg/ml Hoechst 33258 (Sigma-Aldrich, USA). The cells were then rinsed with PBS three times, mounted on slides and examined using a universal fluorescence microscope (Axiophoto, Carl Zeiss, Germany) or confocal laser-scanning microscope (LSM700, Carl Zeiss, Germany). The primary antibodies used in these analyses were as follows: HB9 (mouse IgG1, 1:200, Developmental Studies Hybridoma Bank [DSHB], USA), ISL-1 (mouse IgG2a, 1:200, DSHB, USA), βIII-Tubulin (mouse IgG2b, 1:4000, Sigma-Aldrich, USA or rabbit IgG, 1:4000, Biolegend, USA), ChAT (goat IgG, 1:200, Millipore, USA), GFP (rabbit IgG, 1:200, MBL, USA), and MyHC (mouse IgG2b, 1:200, DSHB, USA). For the observation of n-AChR clusterization, Alexa fluor 555-conjugated α-Bungarotoxin, (1 μg/mL, Invitrogen, USA) was added to samples and incubated for 1 h.
Shimojo D., Onodera K., Doi-Torii Y., Ishihara Y., Hattori C., Miwa Y., Tanaka S., Okada R., Ohyama M., Shoji M., Nakanishi A., Doyu M., Okano H, & Okada Y. (2015). Rapid, efficient, and simple motor neuron differentiation from human pluripotent stem cells. Molecular Brain, 8, 79.
+ Open protocol
+ Expand
3

Claudin Expression and Therapeutic Effect

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 5

CLDN3, 4, 6 and 9 expression was tested by immunoblot analysis in NEC8 cells. The testicular germ cell tumor cell line NEC8 only showed expression of CLDN6 (left panel) but not of CLDN3, 4 or 9, respectively (right panels); see FIG. 15. The specificities of the anti-CLDN3, 4 and 9 antibodies used were tested by Western blot using HEK293T cells transiently transfected with expression vectors encoding for the corresponding human claudin.

CLDN6 surface expression on NEC8 cells was analyzed using flow cytometry. The commercially available anti-CLDN6 antibody (R&D Systems, MAB3656) detected expression of CLDN6 on NEC8 cells; see FIG. 16. Mouse IgG2b (Sigma, CRL M8894) was used as an isotype control.

The therapeutic effect of muMAB 5F2D2 in an early treatment xenograft model using mice engrafted with the tumor cell line NEC8 was tested. Compared to the saline control group muMAB 5F2D2 showed specific and strong tumor growth inhibition in mice engrafted with NEC8 cells that endogenously express human CLDN6; see FIG. 17. The Kruskal-Wallis test shows that tumor volumes were reduced at day 21 and 42 after treatment with muMAB 5F2D2; see FIG. 18.

US10844133B2. Cancer therapy using CLDN6 target-directed antibodies in vivo (2020-11-24). Ganymed Pharmaceuticals AG [DE], Johannes Gutenberg-Universitat Mainz [DE]. Inventors: Ugur Sahin [DE], Ozlem Tureci [DE], Michael Koslowski [DE], Korden Walter [DE], Maria Kreuzberg [DE], Sylvia Luxen [DE].
+ Open protocol
+ Expand
4

Quantification and Characterization of Extracellular Vesicles

Check if the same lab product or an alternative is used in the 5 most similar protocols
MPs were purified from treated cell supernatants as described above. After suspension of MPs in PBS, the MP numbers were quantitated by flow cytometry as described above. For experiments to digest DNA, MPs were treated with recombinant DNase I (rDNase; 10U rDNase / 4×106 MPs; Life Technologies) for 60 min at 37°C. After this treatment, the MPs were transferred to flow cytometry tubes at an MP density of 2×105/tube. To assess HMGB1 content, samples were stained with a Phycoerythrin (PE)-conjugated mouse anti-human HMGB1 antibody (2ul/tube; monoclonal antibody directed to residues 2–215, R&D systems, Minneapolis, MN) or a PE-conjugated mouse IgG2b (R&D Systems) for 60min at 20°C. To assess DNA, MPs were stained with a mouse monoclonal anti-dsDNA antibody (clone 163p.132; 4ug/tube; a kind gift from Dr. Tony Marion) or a mouse IgG2b (isotype control; 4ug/tube; Sigma-Aldrich). After this step, MPs treated with either anti-DNA or isotype control were incubated with a F(ab')2 sheep anti-mouse IgG PE (Sigma-Aldrich) for 30min at 20°C in the dark. The samples were then analyzed by flow cytometry.
Spencer D.M., Mobarrez F., Wallén H, & Pisetsky D.S. (2014). The Expression of HMGB1 on Microparticles from Jurkat and HL-60 Cells Undergoing Apoptosis in vitro. Scandinavian journal of immunology, 80(2), 101-110.
+ Open protocol
+ Expand
5

Quantitative Analysis of β2-Adrenergic Receptor Internalization

Check if the same lab product or an alternative is used in the 5 most similar protocols
After expressing Flag-β2AR, PKA mutant β2AR or GRK mutant β2AR, H9c2 cardiac myoblasts and MEFs were serum-starved for 24h and stimulated with isoproterenol (10 μM, 5min) or insulin (100 nM, 10 min) to examine receptor internalization. Cells were fixed, permeabilized, and stained with anti-flag antibody (mouse IgG2b, Sigma, MO), which was revealed with an Alexa-fluor-488 conjugated goat anti-mouse IgG2b antibody (Life technologies, CA). Fluorescence images were taken with a CCD camera on a Zeiss microscope with Meta morph software (Molecular Devices, CA). Images were quantified with Image J-based Fiji open source image-processing package as described previously [16 (link)]. Briefly, images were rotated to enable optimal selection of areas across the cell body. Plot profile analysis was applied to selected regions for measure of fluorescence intensity. Average of cytosolic fluorescence intensity (CFI) of β2AR staining was divided by that of membrane fluorescence intensity (MFI) for analysis of β2AR internalization. The ratio of CFI/MFI after stimulation was normalized against the ratio of CFI/MFI at resting state.
Fu Q., Xu B., Prakh D., Cervantes D, & Xiang Y.K. (2014). Insulin induces IRS2-dependent and GRK2-mediated β2AR internalization to attenuate βAR signaling in cardiomyocytes. Cellular signalling, 27(3), 707-715.
+ Open protocol
+ Expand
6

Antibody-mediated Cell Cytotoxicity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells in DMEM supplemented with 10% FBS (2x104 cells/well) were plated in 96-well plates., and incubated for 5 h at 37°C with either anti-HER2 antibodies or control IgG (mouse IgG2b) (Sigma-Aldrich Corp.) and 10% of rabbit complement (Low-Tox-M Rabbit Complement) (Cedarlane Laboratories). To assess cell viability, an MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; inner salt] assay was performed using a CellTiter 96 AQueous assay kit (Promega).
Takei J., Kaneko M.K., Ohishi T., Kawada M., Harada H, & Kato Y. (2020). H2Mab-19, an anti-human epidermal growth factor receptor 2 monoclonal antibody exerts antitumor activity in mouse oral cancer xenografts. Experimental and Therapeutic Medicine, 20(2), 846-853.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!