Antibody data
- Antibody Data
- Antigen structure
- References [5]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [6]
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- Product number
- 48-9994-42 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Perforin Monoclonal Antibody (dG9 (delta G9)), eFluor™ 450, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The dG9 antibody clone reacts with human perforin (pore-forming protein, pfp). Perforin is one of the cytolytic mediators present in the cytoplasmic granules of cytotoxic T lymphocytes (CTL) and natural killer cells (NK). Perforin is involved in the killing function by CTLs and NKs and has an important role in the immune response against tumors and virus infections. Applications Reported: This dG9 (delta G9) antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This dG9 (delta G9) antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resource Tab online. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human, Porcine
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- dG9 (delta G9)
- Vial size
- 100 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references LINC01123 promotes immune escape by sponging miR-214-3p to regulate B7-H3 in head and neck squamous-cell carcinoma.
Human Thymic CD10(+) PD-1(+) Intraepithelial Lymphocyte Precursors Acquire Interleukin-15 Responsiveness at the CD1a(-) CD95(+) CD28(-) CCR7(-) Developmental Stage.
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Peritoneal carcinomatosis of colorectal cancer is characterized by structural and functional reorganization of the tumor microenvironment inducing senescence and proliferation arrest in cancer cells.
Lipoxin A4 regulates natural killer cell and type 2 innate lymphoid cell activation in asthma.
Li H, Yang Z, Yang X, Zhang F, Wang J, Wu Z, Wanyan C, Meng Q, Gao W, Yang X, Wei J
Cell death & disease 2022 Feb 3;13(2):109
Cell death & disease 2022 Feb 3;13(2):109
Human Thymic CD10(+) PD-1(+) Intraepithelial Lymphocyte Precursors Acquire Interleukin-15 Responsiveness at the CD1a(-) CD95(+) CD28(-) CCR7(-) Developmental Stage.
Billiet L, Goetgeluk G, Bonte S, De Munter S, De Cock L, Pille M, Ingels J, Jansen H, Weening K, Van Nieuwerburgh F, Kerre T, Taghon T, Leclercq G, Vandekerckhove B
International journal of molecular sciences 2020 Nov 20;21(22)
International journal of molecular sciences 2020 Nov 20;21(22)
Blocking the recruitment of naive CD4(+) T cells reverses immunosuppression in breast cancer.
Su S, Liao J, Liu J, Huang D, He C, Chen F, Yang L, Wu W, Chen J, Lin L, Zeng Y, Ouyang N, Cui X, Yao H, Su F, Huang JD, Lieberman J, Liu Q, Song E
Cell research 2017 Apr;27(4):461-482
Cell research 2017 Apr;27(4):461-482
Peritoneal carcinomatosis of colorectal cancer is characterized by structural and functional reorganization of the tumor microenvironment inducing senescence and proliferation arrest in cancer cells.
Seebauer CT, Brunner S, Glockzin G, Piso P, Ruemmele P, Schlitt HJ, Geissler EK, Fichtner-Feigl S, Kesselring R
Oncoimmunology 2016;5(12):e1242543
Oncoimmunology 2016;5(12):e1242543
Lipoxin A4 regulates natural killer cell and type 2 innate lymphoid cell activation in asthma.
Barnig C, Cernadas M, Dutile S, Liu X, Perrella MA, Kazani S, Wechsler ME, Israel E, Levy BD
Science translational medicine 2013 Feb 27;5(174):174ra26
Science translational medicine 2013 Feb 27;5(174):174ra26
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Staining of normal human peripheral blood cells with Anti-Human CD56 (NCAM) APC (Product # 17-0567-42) and Mouse IgG2b K Isotype Control eFluor® 450 (Product # 48-4732-82) (left) or Anti-Human Perforin eFluor® 450 (right) using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
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- Submitted by
- Invitrogen Antibodies (provider)
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- Submitted by
- Invitrogen Antibodies (provider)
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- Figure 3 Proliferation with IL-15 induces cytotoxic mediators in CD10 + PD-1 + IELps. ( A ) fold expansion of TCRgammadelta + , conventional and IELp cells cultured for 8 days under various conditions (n = 2); ( B ) IFN-gamma, granzyme B and perforin production by unstimulated TCRgammadelta + , conventional and IELp cells incubated for 5 days in the presence of IL-7, IL-15 or IL-15 + IL-12 + IL-18. IFN-gamma, granzyme B, and perforin production measured via intracellular flow cytometry. Bottom: Cytokine production in IELps, plotted for the first and sixth/last generation ( Figure S5 ; mean +- SEM, n = 3); ( C ) flow cytometric analysis of IFN-gamma, granzyme B and perforin production by TCRgammadelta + cells and IELps proliferated for 6 days with IL-15, followed by stimulation with IL-12 + IL-18 for 48 h or with PMA + ionomycin for 4 h (mean +- SEM, n = 3). Dunnett''s multiple comparisons test was used to assess statistically significant differences in cytokine production between the different populations and different interleukins. p -value < 0.05 (*), p < 0.01 (**), and p < 0.001 (***).
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- Invitrogen Antibodies (provider)
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- Fig. 4 Upregulation of LINC01123 or B7-H3 or downregulation of miR-214-3p induces dysfunction of CD8 + T cells. CD8 + T cells were isolated from PBMCs using immunomagnetic beads. After activation and amplification, CD8 + T cells were cocultured with different CAL-27 cell treatment groups. A Expression of TNF-alpha, IFN-gamma, perforin, and granzyme B in CD8 + T-cell population was analyzed with flow cytometry. B Statistical analysis of the results from flow cytometry of CD8 + T cells. C Expression levels of TNF-alpha, IFN-gamma, perforin, and granzyme B in CAL-27-CD8 + T cocultured supernatants, as detected by ELISA. D CCK8 cytotoxicity test of CD8 + T cells. E ELISA was used to assess the immune activity of CD8 + T cells after recombinant human B7-H3 treatment. * P < 0.05 compared with cells without treatment. The experiment was repeated three times.
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- Fig. 6 The function of LINC01123 in HNSCC can be reversed by miR-214-3p silencing. HNSCC cells were transfected with sh-LINC01123 in the presence of inhibitor-NC or miR-214-3p inhibitor. A Relative expression levels of LINC01123, miR-214-3p, and B7-H3 were determined by qRT-PCR. B B7-H3 protein expression was determined by western blot analysis. C CAL-27 cell viability was measured using a CCK8 assay. D Migration of CAL-27 cells was determined by scratch wound-healing assay. E Statistical results of the scratch-healing test. F Migration and invasion of CAL-27 cells, as determined by transwell migration and invasion assay. G Number of migration and invasion cells. H Apoptosis rate and cell-cycle stage were detected by flow cytometry. I Statistical analysis of apoptosis results. J Statistical analysis of cell cycle stage results. K CD8 + T cells were cocultured with CAL-27 cells that had been transfected with plasmids, and the expression of TNF-alpha, IFN-gamma, perforin, and granzyme B in the CD8 + T cells was analyzed by flow cytometry. L Statistical analysis of CD8 + T-cell flow cytometry results. M After different treatments, CAL-27 cells were cocultured with CD8 + T cells, and the expression levels of TNF-alpha, IFN-gamma, perforin, and granzyme B in coculture supernatants were detected by ELISA. N Killing effect of CD8 + T cells on CAL-27 cells, as assessed by CCK8. O Tumor-volume growth curves of the sh-LINC01123 + inhibitor-NC group and sh-LINC01123 + miR-214-3p-inhibit
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- Figure 3 Naive CD4 + T cells are converted to functional Tregs by tumor-infiltrating DCs and tumor conditioned medium (CM). (A-C) Naive CD4 + T cells from peripheral blood of patients with invasive breast carcinoma were co-cultured with or without autologous pDCs isolated from tumor (TI) or peripheral blood (PB) for 9 days in the presence or absence of 30% CM from autologous tumor slices or adjacent normal tissue slices. (A , B) Non-adherent cells from co-cultures were stained for CD3, CD4, CD25 and intracellular Foxp3, and analyzed by flow cytometry. Representative plots of gated CD3 + CD4 + cells (A) and quantification of percentage of Foxp3 + CD25 + cells among CD3 + CD4 + cells (B) are shown (mean +- SEM, n = 19; * P < 0.05, ** P < 0.01, *** P < 0.001 by Student's t -test). (C) Expression of Treg-associated genes, assessed by qRT-PCR normalized to GAPDH , in sorted CD4 + T cells, relative to expression in cultures without DCs or CM (mean +- SEM, n = 19; * P < 0.05, ** P < 0.01, *** P < 0.001 compared with naive CD4 + T cells cultured alone by Student's t -test). (D-G) Effect of naive CD4 + T cell-derived Tregs, obtained by co-culture with TI pDCs and tumor CM as above, on function of autologous tumor-specific CD8 + T cells. Tumor-specific CD8 + T cells were generated for each subject by stimulating autologous PB CD8 + T cells with autologous tumor lysate-pulsed autologous DCs. Tregs were recovered from co-cultures by magnetic sorting. (D) CFSE-labeled CD8 + T ce