Mouse igg2a

Manufactured by Merck Group

Sourced in United States, United Kingdom

Mouse IgG2a is an immunoglobulin G (IgG) isotype produced by mouse B cells. It is a laboratory reagent commonly used in various immunological applications, such as ELISA, Western blotting, and immunohistochemistry, to detect and quantify target proteins or antigens.

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Lab products found in correlation

Rabbit iba1, by Fujifilm (2 mentions) Mouse igg1, by Developmental Studies Hybridoma Bank (2 mentions) Mc5 sc 80, by Synaptic Systems (2 mentions) Mouse igg1, by BD (1 mentions) B27 supplement, by Thermo Fisher Scientific (1 mentions) Glutamax supplement, by Thermo Fisher Scientific (1 mentions) Matrigel, by Corning (1 mentions) Neurobasal medium, by Thermo Fisher Scientific (1 mentions) Ncl ki67p, by Merck Group (1 mentions) T8660, by Merck Group (1 mentions) Sheep serum, by Merck Group (1 mentions) Matched pair antibodies, by Mabtech (1 mentions) Rat anti mouse cd16 cd32 fc block 2.4g2, by BD (1 mentions) Ventana benchmark ultra, by Roche (1 mentions) Clone 3f10, by Roche (1 mentions) Anti flag dykddddk, by Cell Signaling Technology (1 mentions) Clone 9e10, by Santa Cruz Biotechnology (1 mentions) Anti ha, by Roche (1 mentions) Anti myc, by Santa Cruz Biotechnology (1 mentions) Protein a or g agarose beads, by GE Healthcare (1 mentions)

Close spelling variants of this product

IgG2a (35 citations) Anti-mouse IgG1 and IgG2a horseradish-peroxidase conjugated antibodies (7 citations) Goat anti-mouse IgG1 and IgG2a secondary antibodies (4 citations) Anti-mouse IgG2a (3 citations) Mouse IgG2a isotype control (3 citations) FITC-conjugated mouse IgG2a (3 citations) Control mouse IgG2a (3 citations) Anti- αSMA (mouse IgG2a (2 citations) Mouse IgG2a anti-human monoclonal HLA class I antibody clone W6/32 (2 citations) Control IgG (mouse IgG2a) (2 citations)

17 protocols using mouse igg2a

Immunofluorescence Staining of Cochlear Tissues

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For immunofluorescence staining, we used antibodies against rhodamine-phalloidin (1:200, Yeasen, #40734ES75), C-terminal binding protein-2 (CtBP2, 1:400, mouse IgG1, BD Bioscience, #612044¹), glutamate receptor 2 (GluR2, 1:200, mouse IgG2a, Millipore, Cat #mab397²), and 4′, 6-diamidino-2-phenylindole (1:1,000, Sigma, United States, #D9542).
Whole-mount organs of Corti were permeabilized with 0.1% Triton X-100 for 40 min and then blocked with 10% normal goat serum for 30 min at 37°C. Subsequently, the tissue was incubated with the primary antibodies overnight at 4°C. After rinsing, the samples were incubated with their specific secondary antibodies: 1:500 of goat anti-mouse IgG1 (Invitrogen, Alexa Flour™ 568-conjugated, ref#A21134, RRID: AB_2535766) and goat anti-mouse IgG2a (Invitrogen, Alexa Flour™ 647-conjugated, ref#A21241, RRID: AB_2535810) at 37°C for 2 h. The samples were imaged using a laser confocal microscope (Leica SP8, Germany).

Hou S., Zhang J., Wu Y., Junmin C., Yuyu H., He B., Yang Y., Hong Y., Chen J., Yang J, & Li S. (2022). FGF22 deletion causes hidden hearing loss by affecting the function of inner hair cell ribbon synapses. Frontiers in Molecular Neuroscience, 15, 922665.

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Retinal Immunostaining Protocol

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Retina sections were rehydrated in 1XPBS for 10 minutes, followed by incubation in blocking solution: PBS, 5% normal donkey serum, 0.1% triton for 20 minutes at room temperature. Primary antibodies were added to 0.1% block and incubated on sections overnight at 4°C. Sections were washed 3 x 10 minutes, followed by adding secondary antibodies diluted in 0.1% blocking solution and incubated for two hours at room temperature. Finally, a 1:100,000 dilution of DAPI 10 mg/ml in DMSO was added to the second of three final washes in 1XPBS, followed by coverslip mounting in 80% glycerol. Primary antibodies used: protein kinase C alpha (PKCα; anti-mouse, 1:100, Santa Cruz Biotechnology, MC5: sc-80), ribeye (anti-rabbit, 1:500, Synaptic Systems, #192 003), dystroglycan (Mouse IgG₁, Developmental Studies Hybridoma Bank, mandag2 clone 7D11), glutamine synthase (Mouse IgG_2a, 1:2000, Millipore, #MAB302), ionized calcium binding adapter molecule 1 (Iba1; rabbit, 1:500, WAKO Pure Chemical Industries, Ltd., #019–19741). All secondary antibodies were purchased from Jackson ImmunoResearch and used at 1:1000 dilutions.

Clemons M.R., Dimico R.H., Black C., Schlussler M.K., Camerino M.J., Aldinger-Gibson K., Bartle A., Reynolds N., Eisenbrandt D., Rogers A., Andrianu J., Bruce B., Elliot A., Breazeal T., Griffin H., Murphy M.K, & Fuerst P.G. (2023). The rod synapse in aging wildtype and Dscaml1 mutant mice. PLOS ONE, 18(11), e0290257.

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Retinal Immunostaining Protocol

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Neural Stem Cell Differentiation Protocol

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Matrigel (Corning, Corning, NY, USA) and Neurobasal Medium (Thermo Fisher Scientific) containing B27 supplement (Thermo Fisher Scientific) and GlutaMAX supplement (Thermo Fisher Scientific) were mixed at 1:100. Matrix coating was performed using 300 µL of this mixture, and NS/PCs were incubated for a day. The supernatant was then removed, and NS/PCs were incubated for 14 days in Neurobasal Medium containing 0.05 µL of DAPT (Fujifilm) per mL. Half of the medium was changed every 4 days. Subsequently, 5-FC was administered for 6 days. Cells were fluorescently stained with anti-Nestin (1:200, mouse IgG; Chemicon, Tokyo, Japan; MAB5326), anti-Ki67 (1:500, rabbit IgG; Novocastra, Newcastle, UK; NCL-Ki67p), and anti-beta III tubulin (1:200, mouse IgG2a; Sigma-Aldrich; T8660) antibodies.

Imai R., Tamura R., Yo M., Sato M., Fukumura M., Takahara K., Kase Y., Okano H, & Toda M. (2023). Neuroprotective Effects of Genome-Edited Human iPS Cell-Derived Neural Stem/Progenitor Cells on Traumatic Brain Injury. Stem Cells, 41(6), 603-616.

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Unilateral Ureteral Obstruction Induces Renal Fibrosis

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To induce renal fibrosis, wild-type C57BL/6 male mice (Japan SLC) were subjected to unilateral ureteral obstruction (UUO), in which the left ureter was cut between 2 ligatures made by 4–0 Polysorb (Syneture) at the level of the lower pole of the kidney [25] (link). Injured (left) and contralateral (right) kidneys were analysed at 3, 14, and 21 days after UUO surgery. Myofibroblasts were detected in tissue sections from injured kidneys by means of immunofluorescence with Cy3-conjugated anti-human α-smooth muscle actin (αSMA) antibody (mouse IgG2a, Sigma, C6198) [25] (link).

Miyauchi K., Nakai T., Saito S., Yamamoto T., Sato K., Kato K., Nezu M., Miyazaki M., Ito S., Yamamoto M, & Suzuki N. (2021). Renal interstitial fibroblasts coproduce erythropoietin and renin under anaemic conditions. EBioMedicine, 64, 103209.

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Neutralizing Type I Interferon in PBMCs

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To neutralise type I IFNs, PBMCs were treated with reovirus for 16 h±neutralising antibodies (NAbs; IFN block; PBL Interferon Source, Piscataway, NJ, USA) or isotype control (R&D Systems). IFN block consisted of sheep polyclonal antibodies (0.75% of anti-IFN-α and -β) and mouse monoclonal anti-human IFN-α/β receptor chain 2 (1.25%). For isotype controls, 1.5% (v/v) sheep serum (Sigma-Aldrich) and 1.25% (v/v) mouse IgG2a was used. Secretion of IFNα from PBMCs (±reovirus) was determined by enzyme-linked immunosorbent assay (ELISA) using matched-pair antibodies (Mabtech, Cincinnati, OH USA).

Parrish C., Scott G.B., Migneco G., Scott K.J., Steele L.P., Ilett E., West E.J., Hall K., Selby P.J., Buchanan D., Varghese A., Cragg M.S., Coffey M., Hillmen P., Melcher A.A, & Errington-Mais F. (2015). Oncolytic reovirus enhances rituximab-mediated antibody-dependent cellular cytotoxicity against chronic lymphocytic leukaemia. Leukemia, 29(9), 1799-1810.

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Anti-GPC1 Antibody Inhibits Esophageal Cancer Growth

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Example 11

An esophageal squamous cell carcinoma cell strain TE14 was subcutaneously implanted to NOD/SCID mice (6-week old, female) at 2×10⁶cells/100 μl (PBS:Matrigel=1:1). On day 14 after the implantation, the mice were divided into two groups and anti-GPC1 antibody #4(1-12), anti-GPC1 antibody #19(2-70), or the isotype control antibody (Mouse IgG2a, M7769, Sigma) was intraperitoneally administered at 10 mg/kg at a frequency of twice a week and a total of 6 times. The TE14-implanted mice were dissected on day 24 after the start of the antibody administration, and the tumor weight is also measured. It was calculated according to the following formula: tumor volume=major axis×minor axis×minor axis×0.5.

As a result of measuring a tumor volume, a significantly, but partially, inhibitory effect on in vivo growth of tumor in the NOD/SCID mice was recognized in anti-GPC1 antibody #4 (1-12) administered group relative to the control IgG administered group (FIG. 21). A significant difference in tumor weight was also recognized. A similar result was also recognized in tumor weight (FIG. 22).

US10077316B2. Esophageal cancer marker and use thereof (2018-09-18). National University Corporation Kochi University [JP]. Inventors: Tetsuji Naka [JP], Satoshi Serada [JP], Minoru Fujimoto [JP], Masayoshi Toyoura [JP], Yuji Shoya [JP].

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Evaluating Anti-LSR Antibody's Effect on Ovarian Clear Cell Adenocarcinoma

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Example 5

An ovarian clear cell adenomatous cancer cell strain RMG-I was subcutaneously implanted to NOD/Scid mice (6-week old, female) at 1×10⁶cells/100 μl (PBS:Matrigel®=1:1). On day 14 after the implantation, the mice were divided into two groups and an anti-LSR antibody (#1-25) or an isotype control antibody (Mouse IgG2a, M7769, Sigma) was intraperitoneally administered at 10 mg/kg at a frequency of twice a week and a total of 6 times (FIG. 43). The RMG-I-implanted mice were dissected on day 25 after the start of the antibody administration, and the tumor weight was also measured. The following was calculated: tumor volume=major axis×minor axis×height.

As a result of measuring a tumor volume, in the NOD/Scid mice, a significantly inhibitory effect on tumor growth in vivo was exhibited in the anti-LSR antibody administered group relative to the control IgG administered group (FIG. 44). A significant difference in tumor weight was also recognized. In addition, as a result of immunohistochemically staining a tumor tissue with an anti-Ki-67 antibody, a significant decrease in the number of Ki-67 positive cell was recognized in the anti-LSR antibody administered group in comparison with the control IgG administered group. From this, it became clear that the anti-LSR antibody exhibits activity of inducing the arrest of the cell cycle in vivo (FIG. 45).

US10174111B2. Therapeutic drug for malignant tumors (2019-01-08). National University Corporation Kochi University [JP]. Inventors: Tetsuji Naka [JP], Satoshi Serada [JP], Minoru Fujimoto [JP], Masayoshi Toyoura [JP], Yuji Shoya [JP].

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Anti-LSR Antibody Suppresses Ovarian Cancer Growth

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Example 4

An ovarian clear cell adenomatous cancer cell strain RMG-I was subcutaneously implanted to Scid mice (6-week old, female) at 1×10⁶cells/100 μl (PBS:Matrigel®=1:1). On day 14 after the implantation, the mice were divided into two groups and an anti-LSR antibody (#1-25) or an isotype control antibody (Mouse IgG2a, M7769, Sigma) was intraperitoneally administered at 10 mg/kg at a frequency of twice a week and a total of 6 times (FIG. 25). The RMG-I-implanted mice were dissected on day 25 after the start of the antibody administration, and the tumor weight was also measured. The following was calculated: tumor volume=major axis×minor axis×height.

As a result of measuring a tumor volume, a significantly inhibitory effect on tumor growth in vivo was exhibited in the anti-LSR antibody administered group relative to the control IgG administered group (FIGS. 26 to 28). A significant difference in tumor weight was also recognized.

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Fc Receptor Blocking for IgG3 Binding

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IgG3 binding to P388D1 and J774A.1 cells was analyzed in the presence of antibodies that block Fc receptors. FcγRII and FcγRIII were blocked with rat anti-mouse CD16/CD32 Fc-block 2.4G2 (20 µg/ml; BD). FcγRI and FcγRIV were blocked with mouse IgG2a (20, 50 or 100 µg/ml, Sigma, cat. #M5409). The putative non-canonical Fc receptor ITGB1 was blocked with a hamster HMβ1-1 anti-mouse CD29 antibody (20 µg/ml, BD). The cells were incubated with the blocking antibodies for 30 min at RT. The mIgG3 or its derivative was then added directly to the cells with blocking antibodies, followed by the rosetting assay or flow cytometry analysis.

Karabasz A., Bzowska M., Bereta J., Czarnek M., Sochalska M, & Klaus T. (2021). Mouse IgG3 binding to macrophage-like cells is prevented by deglycosylation of the antibody or by Accutase treatment of the cells. Scientific Reports, 11, 10295.

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