Antibody data
- Antibody Data
- Antigen structure
- References [7]
- Comments [0]
- Validations
- Flow cytometry [1]
- Other assay [7]
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Validation data
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- Product number
- 67-0049-41 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CD4 Monoclonal Antibody (RPA-T4), Super Bright™ 702, eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: The RPA-T4 monoclonal antibody reacts with human CD4, a 59 kDa cell surface receptor expressed by a majority of thymocytes, subpopulation of mature T cells (T-helper cells) and in low levels on monocytes. CD4 is a receptor for the human immunodeficiency virus (HIV). RPA-T4 blocks HIV binding and mixed lymphocyte reaction. The RPA-T4 antibody recognizes a different epitope than the OKT4 monoclonal antibody, and these antibodies do not cross-block binding to each other's respective epitopes. Applications Reported: This RPA-T4 antibody has been reported for use in flow cytometric analysis. Applications Tested: This RPA-T4 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. 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. Super Bright 702 is a tandem dye that can be excited with the violet laser line (405 nm) and emits at 702 nm. We recommend using a 710/50 bandpass filter. Please make sure that your instrument is capable of detecting this fluorochrome. When using two or more Super Bright dye-conjugated antibodies in a staining panel, it is recommended to use Super Bright Complete Staining Buffer (Product # SB-4401) to minimize any non-specific polymer interactions. Please refer to the datasheet for Super Bright Staining Buffer for more information. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 405 nm; Emission: 702 nm; Laser: Violet Laser Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- RPA-T4
- Vial size
- 25 Tests
- Concentration
- 5 µL/Test
- Storage
- 4° C, store in dark, DO NOT FREEZE!
Submitted references CD4 + T cells are found within endemic Burkitt lymphoma and modulate Burkitt lymphoma precursor cell viability and expression of pathogenically relevant Epstein-Barr virus genes.
Induced Human Regulatory T Cells Express the Glucagon-like Peptide-1 Receptor.
Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation.
HDAC Inhibitor, CG-745, Enhances the Anti-Cancer Effect of Anti-PD-1 Immune Checkpoint Inhibitor by Modulation of the Immune Microenvironment.
Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells.
CXCL13-producing TFH cells link immune suppression and adaptive memory in human breast cancer.
Spontaneous Chitin Accumulation in Airways and Age-Related Fibrotic Lung Disease.
Sidorov S, Fux L, Steiner K, Bounlom S, Traxel S, Azzi T, Berisha A, Berger C, Bernasconi M, Niggli FK, Perner Y, Pather S, Kempf W, Nadal D, Bürgler S
Cancer immunology, immunotherapy : CII 2022 Jun;71(6):1371-1392
Cancer immunology, immunotherapy : CII 2022 Jun;71(6):1371-1392
Induced Human Regulatory T Cells Express the Glucagon-like Peptide-1 Receptor.
Rode AKO, Buus TB, Mraz V, Al-Jaberi FAH, Lopez DV, Ford SL, Hennen S, Eliasen IP, Klewe IV, Gharehdaghi L, Dragan A, Rosenkilde MM, Woetmann A, Skov L, Ødum N, Bonefeld CM, Kongsbak-Wismann M, Geisler C
Cells 2022 Aug 19;11(16)
Cells 2022 Aug 19;11(16)
Murlentamab, a Low Fucosylated Anti-Müllerian Hormone Type II Receptor (AMHRII) Antibody, Exhibits Anti-Tumor Activity through Tumor-Associated Macrophage Reprogrammation and T Cell Activation.
Prat M, Salon M, Allain T, Dubreuil O, Noël G, Preisser L, Jean B, Cassard L, Lemée F, Tabah-Fish I, Pipy B, Jeannin P, Prost JF, Barret JM, Coste A
Cancers 2021 Apr 13;13(8)
Cancers 2021 Apr 13;13(8)
HDAC Inhibitor, CG-745, Enhances the Anti-Cancer Effect of Anti-PD-1 Immune Checkpoint Inhibitor by Modulation of the Immune Microenvironment.
Kim YD, Park SM, Ha HC, Lee AR, Won H, Cha H, Cho S, Cho JM
Journal of Cancer 2020;11(14):4059-4072
Journal of Cancer 2020;11(14):4059-4072
Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells.
Møller SH, Mellergaard M, Madsen M, Bermejo AV, Jepsen SD, Hansen MH, Høgh RI, Aldana BI, Desler C, Rasmussen LJ, Sustarsic EG, Gerhart-Hines Z, Daskalaki E, Wheelock CE, Hiron TK, Lin D, O'Callaghan CA, Wandall HH, Andresen L, Skov S
Frontiers in immunology 2020;11:1968
Frontiers in immunology 2020;11:1968
CXCL13-producing TFH cells link immune suppression and adaptive memory in human breast cancer.
Gu-Trantien C, Migliori E, Buisseret L, de Wind A, Brohée S, Garaud S, Noël G, Dang Chi VL, Lodewyckx JN, Naveaux C, Duvillier H, Goriely S, Larsimont D, Willard-Gallo K
JCI insight 2017 Jun 2;2(11)
JCI insight 2017 Jun 2;2(11)
Spontaneous Chitin Accumulation in Airways and Age-Related Fibrotic Lung Disease.
Van Dyken SJ, Liang HE, Naikawadi RP, Woodruff PG, Wolters PJ, Erle DJ, Locksley RM
Cell 2017 Apr 20;169(3):497-509.e13
Cell 2017 Apr 20;169(3):497-509.e13
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Normal human peripheral blood cells were stained with CD3 Monoclonal Antibody, eFluor 450 (Product # 48-0036-42) and Mouse IgG1 kappa Isotype Control, Super Bright 702 (Product # 67-4714-80) (left) or CD4 Monoclonal Antibody, Super Bright 702 (right). Cells in the lymphocyte gate were used for analysis.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NULL
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 4 Murlentamab opsonization of SKOV3-R2 + activates an effective anti-tumor T cell immune response. SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control, 3C23K-CHO normally fucosylated or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors unstimulated (M0) or stimulated with M-CSF and IL-10 (TAMs). After 3 days of co-culture, activated T cells coming from the same healthy donor were added in the culture well for 4 more days. ( A ) The CD4 + Th1/Th2 polarization profile, ( B ) the proportion of CD3 + CD4 + CD25 + regulatory T cells and ( C ) the activation of T CD8 + cells were determined by flow cytometry after four days of co-culture. Data shown (boxplots) are the results from two different experiments (performed with two different healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001. p values were determined using one-way ANOVA analysis followed by Tukey''s multiple comparisons test.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 5 Murlentamab/pembrolizumab combination accentuates the anti-tumoral effect of murlentamab monotherapy through the enhancement of T cell activation. ( A - C ) SKOV3-R2 + ovarian tumor cells were labeled with different 3C23K antibodies (3C23K-FcKO control or murlentamab the low fucosylated form) and cultured in the presence of human monocyte-derived macrophages from healthy donors stimulated with M-CSF and IL-10 (TAMs). After 3 days of co-culture, activated T cells coming from the same healthy donor were added in the culture well for 4 more days. Pembrolizumab was added into co-culture wells everyday from day 3 to day 10. ( A ) Opsonized-SKOV3-R2 + cell number was determined by flow cytometry after one and two days of co-culture with TAMs. Data shown (mean +- SEM) are the results from three different experiments (performed with one healthy donors). ** p < 0.01 compared 3C23K-FcKO vs. Murlentamab. # p < 0.05; ## p < 0.01 compared 3C23K-FcKO + anti-PD-1 vs. Murlentamab + anti-PD-1 as determined using one-way ANOVA analysis followed by Dunnett''s multiple comparisons test. ( B , C ) The CD4 + Th1/Th2 polarization profile and the activation of T CD8 + cells were determined by flow cytometry after four days of co-culture. Data shown (mean +- SEM) are the results from three different experiments (performed with one healthy donors). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. p values were determined using one-way ANOVA analysis followed by Tukey''s multiple compa
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- FIGURE 1 MGAT5 knockout increases NKG2DL expression and activates NKG2D in vitro and in vivo . (A) Surface expression of NKG2D ligands and binding of fluorescently labeled L-PHA (MGAT5 modifications) or E-PHA (MGAT3 modifications) on HEK293 wildtype (WT) and HEK293 MGAT5 knockout (KO) cells or isotype control staining (Iso) analyzed by flow cytometry. Data are presented as histograms representative of at least three independent experiments and in bar graphs showing mean fluorescence intensity (MFI). (B) In vitro NKG2D activation measured as GFP expression in NKG2D negative reporter cells (Control) and NKG2D expressing (NKG2D) reporter cells (target cells) co-cultivated with WT or KO cells (effector cells) for 14-16 h at indicated effector:target (E:T) ratios. (C) NKG2D activation in vivo measured on reporter cells as in (B) after activation by WT or KO at a 1:1 ratio in peritoneum of NMRI mice for approximately 18 h. GFP expression in DiD-labeled reporter cells signifies NKG2D activation and is shown as GFP MFI values of cells from four-six mice per group. (D) NKG2D down-modulation was assessed on NK/CD8 + T cells (target cells) after co-cultivation for 2 h with WT or KO cells (effector cells) at indicated effector:target ratios (E:T). NKG2DLs on target cells were blocked with NKG2D-Fc (BL) or unblocked with IgG1-Fc (UN). The graph depicts surface expression of NKG2D presented relative to surface NKG2D expression on target cells alone. (E) MICA surface expression (left) and L
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- Invitrogen Antibodies (provider)
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- Experimental details
- Figure 6 GLP-1R + CD4 + T cells are found in the skin. First quadrant GLP-1R (red), second quadrant CD4 (green), third quadrant DAPI (grey) and fourth quadrant merged stained fluorescent microscopy images of vehicle-exposed (Vehicle) and nickel-exposed (Nickel) skin from patients with allergic contact dermatitis to nickel. Representative images from three patients (P1, P2, P3).
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- Invitrogen Antibodies (provider)
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- Figure 3 CG-745 increases helper T cells, cytotoxic T cells and natural killer T cells, and decreases Treg: (A) hPBMCs were incubated with CG (CG-745) for 36 hours and a subset of hPBMCs was analyzed using the antibodies indicated in the text; (B) hPBMCs were co-cultured with Huh7, Hep3B, HepG2 or PLC/PRF/5 cells for 36 hours with or without CG, and a subset of hPBMCs was analyzed by Attune Nxt (Invitrogen, USA).