Antibody data
- Antibody Data
- Antigen structure
- References [24]
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- Validations
- Flow cytometry [1]
- Other assay [10]
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- Product number
- 12-7049-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IL-4 Monoclonal Antibody (8D4-8), PE, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The 8D4-8 antibody reacts with human interleukin-4 (IL-4), a 15-19 kDa cytokine secreted by Th2 cells. Applications Reported:The 8D4-8 antibody has been reported for use as a capture antibody for human IL-4 ELISA and for intracellular staining for flow cytometric analysis. Applications Tested: Has been pre-titrated and tested by intracellular staining for flow cytometric analysis. This can be used at test size: 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. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Conjugate
- Yellow dye
- Isotype
- IgG
- Antibody clone number
- 8D4-8
- Vial size
- 25 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references Dysregulated Peripheral Invariant Natural Killer T Cells in Plaque Psoriasis Patients.
Cord Blood T Cells Expressing High and Low PKCζ Levels Develop into Cells with a Propensity to Display Th1 and Th9 Cytokine Profiles, Respectively.
Recombinant human IL-37 inhibited endometriosis development in a mouse model through increasing Th1/Th2 ratio by inducing the maturation of dendritic cells.
Standardized 11-color flow cytometry panel for the functional phenotyping of human T regulatory cells.
CD4(+) T(h) immunogenicity of the Ascaris spp. secreted products.
Th2 Biased Immunity With Altered B Cell Profiles in Circulation of Patients With Sporotrichosis Caused by Sporothrix globosa.
Single-cell multi-omics analysis presents the landscape of peripheral blood T-cell subsets in human chronic prostatitis/chronic pelvic pain syndrome.
Platelet-Derived GARP Induces Peripheral Regulatory T Cells-Potential Impact on T Cell Suppression in Patients with Melanoma-Associated Thrombocytosis.
Effect of combined propofol-sevoflurane anesthesia on immune function in pediatric patients with acute lymphoblastic leukemia.
Lactobacillus rhamnosus GG-Derived Soluble Mediators Modulate Adaptive Immune Cells.
Functionalization-dependent effects of cellulose nanofibrils on tolerogenic mechanisms of human dendritic cells.
CD4 T cell loss and Th2 and Th17 bias are associated with the severity of severe fever with thrombocytopenia syndrome (SFTS).
Genetic Architecture of Adaptive Immune System Identifies Key Immune Regulators.
Impact of Enhancer of Zeste Homolog 2 on T Helper Cell-Mediated Allergic Rhinitis.
Mucosal and systemic immune modulation by Trichuris trichiura in a self-infected individual.
CD16(+) Monocyte Subset Was Enriched and Functionally Exacerbated in Driving T-Cell Activation and B-Cell Response in Systemic Lupus Erythematosus.
A pilot study showing associations between frequency of CD4(+) memory cell subsets at diagnosis and duration of partial remission in type 1 diabetes.
Effects of transcutaneous acupoint electrical stimulation on the imbalance of Th(1), Th(2), Th(17) and T(reg) cells following thoracotomy of patients with lung cancer.
T-helper 17 cell polarization in pulmonary arterial hypertension.
Do patients with a failed metal-on-metal hip implant with a pseudotumor present differences in their peripheral blood lymphocyte subpopulations?
A SnoRNA-derived piRNA interacts with human interleukin-4 pre-mRNA and induces its decay in nuclear exosomes.
Depressive symptoms post hip fracture in older adults are associated with phenotypic and functional alterations in T cells.
BET bromodomain inhibition suppresses TH17-mediated pathology.
Brefeldin A, but not monensin, enables flow cytometric detection of interleukin-4 within peripheral T cells responding to ex vivo stimulation with Chlamydia trachomatis.
Hu Y, Chen Y, Chen Z, Zhang X, Guo C, Yu Z, Xu P, Sun L, Zhou X, Gong Y, Yu Q, Shi Y
Frontiers in cell and developmental biology 2021;9:799560
Frontiers in cell and developmental biology 2021;9:799560
Cord Blood T Cells Expressing High and Low PKCζ Levels Develop into Cells with a Propensity to Display Th1 and Th9 Cytokine Profiles, Respectively.
Perveen K, Quach A, McPhee A, Prescott SL, Barry SC, Hii CS, Ferrante A
International journal of molecular sciences 2021 May 5;22(9)
International journal of molecular sciences 2021 May 5;22(9)
Recombinant human IL-37 inhibited endometriosis development in a mouse model through increasing Th1/Th2 ratio by inducing the maturation of dendritic cells.
Li L, Liao Z, Ye M, Jiang J
Reproductive biology and endocrinology : RB&E 2021 Aug 24;19(1):128
Reproductive biology and endocrinology : RB&E 2021 Aug 24;19(1):128
Standardized 11-color flow cytometry panel for the functional phenotyping of human T regulatory cells.
Manuszak C, Brainard M, Thrash E, Hodi FS, Severgnini M
Journal of biological methods 2020;7(2):e131
Journal of biological methods 2020;7(2):e131
CD4(+) T(h) immunogenicity of the Ascaris spp. secreted products.
Ebner F, Morrison E, Bertazzon M, Midha A, Hartmann S, Freund C, Álvaro-Benito M
NPJ vaccines 2020;5:25
NPJ vaccines 2020;5:25
Th2 Biased Immunity With Altered B Cell Profiles in Circulation of Patients With Sporotrichosis Caused by Sporothrix globosa.
Zu J, Yao L, Song Y, Cui Y, Guan M, Chen R, Zhen Y, Li S
Frontiers in immunology 2020;11:570888
Frontiers in immunology 2020;11:570888
Single-cell multi-omics analysis presents the landscape of peripheral blood T-cell subsets in human chronic prostatitis/chronic pelvic pain syndrome.
Zhang M, Liu Y, Chen J, Chen L, Meng J, Yang C, Yin S, Zhang X, Zhang L, Hao Z, Chen X, Liang C
Journal of cellular and molecular medicine 2020 Dec;24(23):14099-14109
Journal of cellular and molecular medicine 2020 Dec;24(23):14099-14109
Platelet-Derived GARP Induces Peripheral Regulatory T Cells-Potential Impact on T Cell Suppression in Patients with Melanoma-Associated Thrombocytosis.
Zimmer N, Krebs FK, Zimmer S, Mitzel-Rink H, Kumm EJ, Jurk K, Grabbe S, Loquai C, Tuettenberg A
Cancers 2020 Dec 5;12(12)
Cancers 2020 Dec 5;12(12)
Effect of combined propofol-sevoflurane anesthesia on immune function in pediatric patients with acute lymphoblastic leukemia.
Di N, Guo Y, Ding N
Oncology letters 2019 Jul;18(1):35-42
Oncology letters 2019 Jul;18(1):35-42
Lactobacillus rhamnosus GG-Derived Soluble Mediators Modulate Adaptive Immune Cells.
Ludwig IS, Broere F, Manurung S, Lambers TT, van der Zee R, van Eden W
Frontiers in immunology 2018;9:1546
Frontiers in immunology 2018;9:1546
Functionalization-dependent effects of cellulose nanofibrils on tolerogenic mechanisms of human dendritic cells.
Tomić S, Ilić N, Kokol V, Gruden-Movsesijan A, Mihajlović D, Bekić M, Sofronić-Milosavljević L, Čolić M, Vučević D
International journal of nanomedicine 2018;13:6941-6960
International journal of nanomedicine 2018;13:6941-6960
CD4 T cell loss and Th2 and Th17 bias are associated with the severity of severe fever with thrombocytopenia syndrome (SFTS).
Li MM, Zhang WJ, Weng XF, Li MY, Liu J, Xiong Y, Xiong SE, Zou CC, Wang H, Lu MJ, Yang DL, Peng C, Zheng X
Clinical immunology (Orlando, Fla.) 2018 Oct;195:8-17
Clinical immunology (Orlando, Fla.) 2018 Oct;195:8-17
Genetic Architecture of Adaptive Immune System Identifies Key Immune Regulators.
Lagou V, Garcia-Perez JE, Smets I, Van Horebeek L, Vandebergh M, Chen L, Mallants K, Prezzemolo T, Hilven K, Humblet-Baron S, Moisse M, Van Damme P, Boeckxstaens G, Bowness P, Dubois B, Dooley J, Liston A, Goris A
Cell reports 2018 Oct 16;25(3):798-810.e6
Cell reports 2018 Oct 16;25(3):798-810.e6
Impact of Enhancer of Zeste Homolog 2 on T Helper Cell-Mediated Allergic Rhinitis.
Hou TY, Chen MR, Chou YC, Kan PC, Tsai YT, Cha TL
Frontiers in immunology 2017;8:790
Frontiers in immunology 2017;8:790
Mucosal and systemic immune modulation by Trichuris trichiura in a self-infected individual.
Dige A, Rasmussen TK, Nejsum P, Hagemann-Madsen R, Williams AR, Agnholt J, Dahlerup JF, Hvas CL
Parasite immunology 2017 Jan;39(1)
Parasite immunology 2017 Jan;39(1)
CD16(+) Monocyte Subset Was Enriched and Functionally Exacerbated in Driving T-Cell Activation and B-Cell Response in Systemic Lupus Erythematosus.
Zhu H, Hu F, Sun X, Zhang X, Zhu L, Liu X, Li X, Xu L, Shi L, Gan Y, Su Y
Frontiers in immunology 2016;7:512
Frontiers in immunology 2016;7:512
A pilot study showing associations between frequency of CD4(+) memory cell subsets at diagnosis and duration of partial remission in type 1 diabetes.
Moya R, Robertson HK, Payne D, Narsale A, Koziol J, Type 1 Diabetes TrialNet Study Group, Davies JD
Clinical immunology (Orlando, Fla.) 2016 May;166-167:72-80
Clinical immunology (Orlando, Fla.) 2016 May;166-167:72-80
Effects of transcutaneous acupoint electrical stimulation on the imbalance of Th(1), Th(2), Th(17) and T(reg) cells following thoracotomy of patients with lung cancer.
Wu H, Wang K, Li G, Meng D, Han J, Wang G, Li YU
Experimental and therapeutic medicine 2016 Feb;11(2):495-502
Experimental and therapeutic medicine 2016 Feb;11(2):495-502
T-helper 17 cell polarization in pulmonary arterial hypertension.
Hautefort A, Girerd B, Montani D, Cohen-Kaminsky S, Price L, Lambrecht BN, Humbert M, Perros F
Chest 2015 Jun;147(6):1610-1620
Chest 2015 Jun;147(6):1610-1620
Do patients with a failed metal-on-metal hip implant with a pseudotumor present differences in their peripheral blood lymphocyte subpopulations?
Catelas I, Lehoux EA, Hurda I, Baskey SJ, Gala L, Foster R, Kim PR, Beaulé PE
Clinical orthopaedics and related research 2015 Dec;473(12):3903-14
Clinical orthopaedics and related research 2015 Dec;473(12):3903-14
A SnoRNA-derived piRNA interacts with human interleukin-4 pre-mRNA and induces its decay in nuclear exosomes.
Zhong F, Zhou N, Wu K, Guo Y, Tan W, Zhang H, Zhang X, Geng G, Pan T, Luo H, Zhang Y, Xu Z, Liu J, Liu B, Gao W, Liu C, Ren L, Li J, Zhou J, Zhang H
Nucleic acids research 2015 Dec 2;43(21):10474-91
Nucleic acids research 2015 Dec 2;43(21):10474-91
Depressive symptoms post hip fracture in older adults are associated with phenotypic and functional alterations in T cells.
Duggal NA, Upton J, Phillips AC, Hampson P, Lord JM
Immunity & ageing : I & A 2014;11(1):25
Immunity & ageing : I & A 2014;11(1):25
BET bromodomain inhibition suppresses TH17-mediated pathology.
Mele DA, Salmeron A, Ghosh S, Huang HR, Bryant BM, Lora JM
The Journal of experimental medicine 2013 Oct 21;210(11):2181-90
The Journal of experimental medicine 2013 Oct 21;210(11):2181-90
Brefeldin A, but not monensin, enables flow cytometric detection of interleukin-4 within peripheral T cells responding to ex vivo stimulation with Chlamydia trachomatis.
Vicetti Miguel RD, Maryak SA, Cherpes TL
Journal of immunological methods 2012 Oct 31;384(1-2):191-5
Journal of immunological methods 2012 Oct 31;384(1-2):191-5
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Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- Normal human peripheral blood cells were stimulated with PMA and Ionomycin in the presence of brefeldin A for 6 hours. The cells were surface stained with Anti-Human CD3 FITC (Product # 11-0038-42) and intracellularly stained with Anti-Human IL-4 PE (right panel). Left panel demonstrates unlabelled antibody blocking control.
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Supportive validation
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- Figure 6. piR30840 regulates the development of Th2 lymphocytes. ( A and B ) Human CD4 naive cells were cultured in a conditioned medium for Th2 development and then transfected with an s30840 (A) or piR30840 inhibitor (B). IL-4 expression was detected by qRT-PCR and western blotting analyses. P < 0.01. The data represents three independent experiments. ( C ) Detection of IL-4 expression in human PBMCs isolated from humanized NOG mice. IL-4 expression was detected by qRT-PCR analysis. The data was expressed as the mean of three independent experiments. Wilcoxon test was used to calculate the P -value. P < 0.05. ( D ) CD4 naive cells were cultured in the conditioned medium for Th2 development and transfected with s30840 or control. After 7 days, the Th2 T-lymphocytes (intracellular stained as IL-4 + IFN-gamma - ) were detected with FACS analysis. The data was shown as a mean of six independent experiments. P < 0.01. ( E ) The CD4 naive cells were cultured in a conditioned medium for Th2 development with or without the transfection of inhibitor. We adoptively transferred the naive cells into the irradiated NOG mice which were first humanized. After 7 days, Th2 T-lymphocytes (intracellularly stained as IL-4 + IFNgamma - ) were detected by using FACS analysis. The data were expressed as the mean of six independent experiments. Statistical significance between two samples was determined by using the student's t -test. P < 0.01.
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- Figure 5 Blockade of transforming growth factor (TGF)-beta I-III did in part prevent regulatory T cells (Treg) induction. ( A ) CFSE-labeled CD4 + CD25 - T cells were cocultured with platelets in the ratio of 1:50 and were stimulated with anti-CD3 mAb (0.5 ug/mL) and anti-CD28 mAb (1.0 ug/mL) in the presence of either anti-TGF-beta I-III (10 ug/mL) or anti-TGF-beta receptor II (10 ug/mL) antibodies. Antibodies were added at day 0. The expression of Foxp3 and GARP and cell proliferation were determined on day 3 via flow cytometry. ( B ) Production of IL-2 and IFN-gamma was assessed by intracellular flow cytometry on day 6. The graphs show cells cultured in the presence of platelets normalized to CD4 + CD25 - T cells without platelets. Dot plots show one representative result of 10 independent experiments ( n = 10, box and whiskers, medians +- min/max, * p < 0.05, ** p
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- Figure 6 Combining blockade of TGF-beta signaling and GARP led to a complete inhibition of platelet effects. ( A ) CFSE-labeled CD4 + CD25 - T cells were cocultured with platelets in the ratio of 1:50 and were stimulated with anti-CD3 mAb (0.5 ug/mL) and anti-CD28 mAb (1.0 ug/mL). CD4 + CD25 - T cells were incubated for 15 min with TGF-beta receptor II (10 ug/mL) antibody prior to coculture, as indicated. Excess antibody was removed. Pre-treated CD4 + CD25 - T cells were cultured in the presence of either anti-TGF-beta I-III (10 ug/mL) and/or anti-GARP Ab (10 ug/mL) antibodies. Antibodies were added at day 0. The expression of Foxp3, GARP and cell proliferation were determined on day 3 via flow cytometry. ( B ) Production of IL-2 and IFN-gamma was assessed by intracellular flow cytometry on day 6. The graphs show cells cultured in the presence of platelets normalized to CD4 + CD25 - T cells without platelets. Dot plots show 1 representative result of 10 independent experiments ( n = 3, means +- SD, * p < 0.05, ** p
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- Figure 7 Platelet-conditioned medium (PCM) inhibited IFN-gamma production, but failed to induce a Treg phenotype. ( A ) CD4 + CD25 - T cells were cultured in X-Vivo 15 (Lonza, Basel, Switzerland) with 60% PCM content, with or without 10 ug/mL anti-GARP Ab and stimulated with 0.5 ug/mL anti-CD3 mAb and 1.0 ug/mL anti-CD28 mAb. Antibodies were added at day 0. The expression of Foxp3, GARP and cell proliferation were determined at day 3 with flow cytometry. ( B ) Cytokine production of IL-2 and IFN-gamma was measured by intracellular flow cytometry on day 6. Dot plots show one representative result of five independent experiments ( n = 5, box and whiskers, medians +- min/max, * p < 0.05, ** p
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- Figure A4 Thrombin-activated platelets induced a regulatory phenotype in CD4 + CD25 - T cells. 1 x 10 6 CD4 + CD25 - T cells were stimulated with 0.5 ug/mL anti-CD3 mAb and 1.0 ug/mL anti-CD28 mAb with or without 50 x 10 6 platelets for 6 days and treated with or without 10 U/mL thrombin. ( A ) Foxp3 and GARP expression and proliferation were determined at day 3 via flow cytometry. ( B ) Using intracellular flow cytometry, we analyzed cytokine production of IL-2 and IFN-gamma on day 6. Dot plots show one representative result of five independent experiments ( n = 5, box and whiskers, medians +- min/max, * p < 0.05, ** p
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- Figure A5 TRAP-6-activated platelets induced a regulatory phenotype in CD4 + CD25 - T cells. 1 x 10 6 CD4 + CD25 - T cells were stimulated with 0.5 ug/mL anti-CD3 mAb and 1.0 ug/mL anti-CD28 mAb with or without 50 x 10 6 platelets for 6 days and treated with or without 5 uM TRAP-6. ( A ) Foxp3 and GARP expression and proliferation were determined at day 3 via flow cytometry. ( B ) Using intracellular flow cytometry, we analyzed cytokine production of IL-2 and IFN-gamma on day 6. Dot plots show one representative result of five independent experiments ( n = 5, box and whiskers, medians +- min/max, * p < 0.05, ** p
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- Figure 1 Comparison of proliferation and cytokine production in naive cord blood T cells (CBTC) in Protein Kinase C zeta (PKCzeta) low or high group. ( a ) Comparison of PKCzeta between cord blood (CB) and adult blood (AB) T cells, n = 24 for each AB or CB. ( b ) Shows lymphoproliferation as a Stimulation index (SI) and disintegrations per minute (DPM) in 3 H-thymidine pulsed cultures stimulated with Phytohaemagglutinin (PHA) and Phorbol myristate acetate (PMA) ( c ) Purified CBCTs were stained with Carboxyfluorescein succinimidyl ester (CFSE) dye and stimulated with immobilized anti-CD3/-CD28 antibodies for 3 days. Gating and representative histogram for CFSE dilution after exclusion of doublets and dead cells. Overlaid histograms for stained unstimulated and unstained stimulated samples were used as control and for gating the non-proliferating cells and for auto-fluorescence, respectively. ( d ) Naive CB CD3 + T cells were stimulated with PHA/PMA (18 h) and percentage of CD3 + T cells producing interleukin-4 (IL-4) and Interferon-gamma (IFN-gamma) and median fluorescent intensity (MFI) were examined by flow cytometry assays. ( e ) On day 5 of CFSE stained culture (anti-CD3/-CD28), cells were re-stimulated with PHA/PMA (18 h) for detection of intracellular cytokine. Representative flow dot plots and data for CFSE dye dilution and IFN-gamma producing cells in high and low PKCzeta group. Data mean +- SD of n = 3 for of low and n = 4 high PKCzeta group. ** p < 0.01. ns: not sig
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- Fig. 3 Changes in the percentages and absolute numbers of Th1 and Th2 cells in SFTS patients. (A): Th1 cells (CD4 + IFN-gamma+) and Th2 cells (CD4 + IL-4+) by flow cytometry in the healthy controls, surviving SFTS patients, and deceased SFTS patients, as defined by flow cytometry. The cells were gated on the CD3 + CD4+ population within the single-cell lymphocyte gate. (B), (D): The percentages and numbers of Th1 and Th2 cells in the healthy controls (n = 11) and the surviving patients with SFTS in the acute phase (n = 30) and SFTS in the recovery phase (n = 30). (C), (E): The percentages and numbers of Th1 and Th2 cells at admission in the surviving patients (n = 30) and the deceased patients (n = 12). (F), (G): Dynamic changes in the percentages and numbers of Th1 and Th2 cells in the surviving patients (n = 30) and the deceased patients (n = 12). These parameters were monitored at indicated time points for the entire hospital stay of the patients, and the dashed line represents the median of the uninfected controls. The data are shown as the median +- 95% CI. Statistical analysis was performed using the Mann-Whitney U test or the Wilcoxon matched pair test. The level of significance is indicated as follows: ns, not significant; *p < .05; **p < .01; ***p < .001; ****p < .0001; Fig. 3
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- Figure 4 CD16 + monocytes promoted T-cell-mediated inflammation in SLE . CD16 + or CD16 - monocytes were cocultured with CD4 + T cells isolated from freshly collected SLE blood or blood bank collected HC blood buffy coat for 5 days in the presence of anti-CD3 (1 mug/mL) and anti-CD28 (1 mug/mL) antibodies and M-CSF (50 ng/mL). Intracellular IFN-gamma, IL-4, and IL-17A expression was detected by flow cytometry after PMA (50 ng/mL), ionomycin (1 mug/mL), and BFA (10 mug/mL) stimulation for 5 h on day 5. The percentage of Treg (CD4 + Foxp3 + ) was also analyzed. Representative pseudocolor dots depicted Th1 (A) , Th2 (B) , Th17 (C) , and Treg (D) frequencies in CD4 + T cells after coculture with each monocyte subset from one patient and one control donor. The proportion of Th1, Th2, Th17, and Treg cells was calculated after coculture of CD4 + T cells with each monocyte. The percentage increases in T-cell subsets in cocultures of monocytes and T cells compared with CD4 + T cells cultured alone were compared between 7 healthy individuals and 10 patients with SLE. Data were expressed as mean +- SD and analyzed by non-parametric paired t test and Mann-Whitney U test. * P < 0.05, ** P < 0.01; NS, no significance.
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