67-0049-42

antibody from Invitrogen Antibodies

Targeting: CD4

Provider product page for 67-0049-42

Flow cytometry

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Antibody data

Antibody Data
Antigen structure
References [7]
Comments [0]
Validations
Flow cytometry [1]
Other assay [7]

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Product number: 67-0049-42 - 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: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

CD4 protein structure - 67-0049-42 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 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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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

Supportive validation

Submitted by: 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).

Supportive validation

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