Goat anti hamster igg

Manufactured by Jackson ImmunoResearch

Goat anti-hamster IgG is a secondary antibody produced in goats and purified to specifically recognize and bind to hamster immunoglobulin G (IgG). It is designed for use in various immunological techniques, such as ELISA, Western blotting, and immunohistochemistry, to detect and visualize hamster IgG in samples.

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Lab products found in correlation

Nupage 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions) Rpmi medium, by Welgene (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Welgene (1 mentions) Goat sera, by Jackson ImmunoResearch (1 mentions) Bovine serum albumin (bsa), by Jackson ImmunoResearch (1 mentions) Q31 378, by BD (1 mentions) Anti pe antibody, by Novus Biologicals (1 mentions) Goat anti rabbit af555, by Thermo Fisher Scientific (1 mentions) R17 809, by BD (1 mentions) Goat anti mouse igg1, by Southern Biotech (1 mentions) Teceleukin, by Roche (1 mentions)

Close spelling variants of this product

Hamster IgG (6 citations) Goat anti–Armenian hamster IgG (5 citations) Alexa Fluor 594 goat anti-hamster IgG secondary antibody (3 citations) Goat anti-Armenian hamster IgG antibodies (3 citations) Peroxidase AffiniPure Goat Anti-Syrian Hamster IgG (H + L) (2 citations) Peroxidase AffiniPure goat anti-Armenian hamster IgG (H+L) (2 citations) Cy3-conjugated goat anti-Armenian hamster IgG (2 citations) Goat anti-Armenian hamster IgG Alexa Fluor 594 (2 citations) Alexa Fluor 647-conjugated goat anti-American-hamster IgG (2 citations) FITC -conjugated goat anti-hamster and cy3- conjugated anti-rabbit IgG (2 citations)

5 protocols using goat anti hamster igg

T cell Stimulation and Signaling Pathway Analysis

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Purified CD4⁺ T cells (2-5×10⁶) were incubated with anti-CD3 (2C11 at 10 μg/ml) alone or in combination with anti-CD28 (37.51 at 10 μg/ml) on ice for 30 min. Cells were washed once in ice-cold RPMI, incubated 2 min at 37°C and then stimulated with goat anti-hamster IgG (Jackson ImmunoResearch Laboratory) for the indicated times. Cells were lysed in ice-cold lysis buffer (50 mM HEPES, 150 mM NaCl, 10 mM NaF, 10 mM iodoacetamide, 1% NP-40, 1 mM PMSF and protease inhibitors) and mechanically sheared using a needle and syringe. Proteins were resolved on NuPage 4-12% BisTris gels (Invitrogen), transferred to PVDF membranes and probed with the indicated antibodies.
For small G proteins analysis, lysates were incubated with the appropriate GST fusion protein (GST-PAK-CRIB, GST-Rhotekin-RBD and GST-Ral-RBD), coupled to glutathione-Sepharose beads for 2h at 4°C. Pull-downs were washed three times in ice-cold lysis buffer, and resolved as described above.

Garçon F, & Okkenhaug K. (2016). PI3Kδ promotes CD4+ T-cell interactions with antigen-presenting cells by increasing LFA-1 binding to ICAM-1. Immunology and Cell Biology, 94(5), 486-495.

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TCR Stimulation and ERK Phosphorylation in Thymocytes

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Total thymocytes or sorted DP cells were rested for 30 min in Roswell Park Memorial Institute (RPMI) medium (Welgene) at 37 °C. Cells were then washed with T-cell medium (TCM, RPMI supplemented with 10% FBS [Welgene], 1% penicillin/streptomycin [Gibco], and 0.1% β-mercaptoethanol [Gibco]) and incubated with biotin-conjugated anti-CD3 (60 μg/ml, 145–2 C11) and anti-CD4 (60 μg/ml, GK1.5) for 20 min on ice. Cells were washed and incubated for 5 min on ice with streptavidin (60 μg/ml, SouthernBiotech) for cross-linking and incubated for the indicated time at 37 °C. To analyze p-ERK levels in virus-transduced thymocytes by flow cytometry, unconjugated anti-CD3 (10 μg/ml, 145–2 C11) and goat anti-hamster IgG (25 μg/ml, Jackson Immunoresearch) were used to stimulate TCR. The cells were incubated for 2 min at 37 °C for TCR stimulation. One milliliter of Cold PBS was added at the end of stimulation and cell pellets were lysed for western blotting or further stained with anti-p-ERK antibody for flow cytometry.

Kim S., Park G.Y., Park J.S., Park J., Hong H, & Lee Y. (2021). Regulation of positive and negative selection and TCR signaling during thymic T cell development by capicua. eLife, 10, e71769.

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Multiparametric Immunofluorescence Staining

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For CD1d tetramer immunofluorescence, fresh thymic lobes, whole lymph nodes or 1–2 mm thickness spleen trans-sections were overnight incubated in 50μl phosphate buffered saline (PBS) solution containing 1:10~1:25 dilution of R-Phycoerythrin (PE) labeled PBS-57 loaded CD1d tetramer in 2% fetal calf serum containing PBS at 4°C. Next day, tissues were washed three times with PBS, fixed with 4% paraformaldehyde (PFA) for one hour and snap frozen. Five micrometer tissue sections were blocked with 5% bovine serum albumin and goat sera (Jackson Laboratory) for 1 hour at 25°C and stained with anti-PE antibody (Novus Biologicals, rabbit polyclonal) followed by goat anti-rabbit AF555 (Life Technology). For human CD2 immunofluorescence, tyramide based amplification was done according to manufacturer’s instruction (PerkinElmer) after staining with anti-human CD2 (RPA2) antibody. RORγt (RORg2, Millipore or Q31-378, BD) and PLZF (D9, SantaCruz or R17-809, BD) were stained followed by goat anti-hamster IgG (Jackson Laboratory) and goat anti-mouse IgG1 (Southern Biotech).

Lee Y.J., Wang H., Starrett G.J., Phuong V., Jameson S.C, & Hogquist K.A. (2015). Tissue specific distribution of iNKT cells impacts their cytokine response. Immunity, 43(3), 566-578.

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CD8+ T Cell Activation Assay

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Goat anti-hamster IgG (Jackson ImmunoResearch) and hCEA were pre-coated (5 μg/ml) overnight at 4°C in 96-well plates. After blocking, 1 μg/ml anti-CD3ε mAb (clone 145-2C11; Immunostep, Salamanca, Spain) was added and incubated at 37°C for 1 h. Purified CD8a+ T cells (CD8a+T Cell Isolation Kit, mouse, Miltenyi Biotec, GmbH) from spleens of C57BL/6 mice were added (2 × 10⁵/well) in complete RPMI + 50 μM 2-mercaptoethanol with purified antibodies at 6.67 nM. As a control, purified mouse CD8a⁺ T cells were cultured alone with the immobilized anti-CD3ε mAb. After 72 h, cell proliferation was assessed using the CellTiter-Glo luminescent assay, and supernatants were collected and assayed for IFNγ secretion by ELISA (Diaclone, Besançon, France). Results are expressed as a mean ± SD (n = 3) from 1 of at least 3 separate experiments.

Mikkelsen K., Harwood S.L., Compte M., Merino N., Mølgaard K., Lykkemark S., Alvarez-Mendez A., Blanco F.J, & Álvarez-Vallina L. (2019). Carcinoembryonic Antigen (CEA)-Specific 4-1BB-Costimulation Induced by CEA-Targeted 4-1BB-Agonistic Trimerbodies. Frontiers in Immunology, 10, 1791.

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Activation and Transduction of Naïve CD8 T Cells

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FACS-purified CD44^lo CD62L^hi naïve CD8 T cells (1×10⁶/well of 24 well plate) from CD45.1 C57BL/6 mice were activated in vitro with soluble 0.1μg/ml anti-CD3 (145-2C11) and 1μg/ml anti-CD28 (37.51) antibodies in plates coated with 18 μg/ml goat anti-hamster IgG (Jackson ImmunoResearch #127-005-099) in the presence of 1×10³ IU hIL-2. Twenty seven hrs after stimulation, cells were spinfected (37°C 1300g for 2h) with the indicated pMHCII-CAR RVs (1ml viral supernatant per each well) and subsequently cultured with 1×10³ IU/ml hIL-2 (Teceleukin, Hoffmann-La Roche) before use. Cells cultured for 2 additional days were used for depletion assays of target TCR Tg cells in vivo. Longer cultures for 4–6 additional days were conducted for EAE experiments. CAR T cell transfer was performed into mice without preconditioning by lymphodepletion or irradiation.

Yi J., Miller A.T., Archambault A.S., Jones A.J., Bradstreet T.R., Bandla S., Hsu Y.S., Edelson B.T., Zhou Y.W., Fremont D.H., Egawa T., Singh N., Wu G.F, & Hsieh C.S. (2022). Antigen-specific depletion of CD4+ T cells by CAR T cells reveals distinct roles of higher and lower affinity TCRs during autoimmunity. Science immunology, 7(76), eabo0777.

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