48-9994-42

antibody from Invitrogen Antibodies

Targeting: PRF1 HPLH2, P1, PFP

Provider product page for 48-9994-42

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [5]
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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!

PRF1 protein structure - 48-9994-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 (***).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 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