16-1529-82

antibody from Invitrogen Antibodies

Targeting: CTLA4 CD, CD152, CELIAC3, CTLA-4, GSE, IDDM12

Provider product page for 16-1529-82

Flow cytometry

Blocking/Neutralizing

Other assay

Antibody data

Antibody Data
Antigen structure
References [13]
Comments [0]
Validations
Other assay [8]

Submit

Product number: 16-1529-82 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD152 (CTLA-4) Monoclonal Antibody (14D3), Functional Grade, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The 14D3 monoclonal antibody reacts with human CD152, also known as cytotoxic T lymphocyte antigen-4 (CTLA-4). CTLA-4, a protein with structural similarities to CD28, is expressed on activated T cells (activated B cells may also express CTLA-4) and binds the B7 family members, CD80 (B7-1) and CD86 (B7-2), with higher affinity than CD28 does. CTLA-4 and CD28 appear to deliver opposing signals to T cells: while CD28 is a potent costimulator, CTLA-4 restricts the progression of T cells to an activated state by inhibiting IL-2 secretion and cellular proliferation. The cytoplasmic portion of CTLA-4 contains ER retention motifs, resulting in intracellular localization of a large proportion of newly synthesized CTLA-4 in response to TCR signaling. The 14D3 antibody also recognizes rhesus monkey and has inhibitor activity. Applications Reported: The 14D3 antibody has been reported for use in flow cytometric analysis. It has also been reported in in vitro inhibition studies. (Please use Functional Grade purified 14D3 in functional assays). Applications Tested: The 14D3 antibody has been tested by flow cytometric analysis of PHA stimulated normal human peripheral blood cells. This can be used at less than or equal to 0.25 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Furthermore, due to the intracellular localization of a large portion of CTLA-4, for complete detection it may be necessary to assess intracellular expression, in addition to surface expression of CTLA-4. Storage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: 14D3
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: 4° C

CTLA4 protein structure - 16-1529-82 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
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 6 T FR exhibit an enhanced regulatory phenotype in ex vivo HIV infection. Tonsil cells were mock-, X4-, or R5-spinoculated and cultured under experimental conditions as indicated. T FR were then analysed for expression of regulatory receptors and cytokine production by intracellular cytokine staining. ( a ) Percentage of total (surface and intracellular) T FR CTLA-4 expression ( n =15). ( b ) Percentage of surface T FR LAG-3 expression ( n =8). ( c ) Production of IL-10 by T FR ( n =7). ( d ) Production of TGF-beta-1 (measured as LAP) by T FR ( n =5). The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Friedman ( a , b ) or Mann-Whitney tests ( c , d ) and significance is denoted by asterisks where * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Resident memory T (T RM ) and non-T RM phenotypic profiles and susceptibility of cancer stem cell (CSC) to cytotoxic T lymphocyte (CTL)-mediated killing. (A) Flow cytometry analyses of CD8, LFA-1, CD103, CD49a, CD69, PD-1, CTLA-4, TGFBR2 and VEGFR2 on CD8 + CD103 + T RM (Heu171) and CD8 + CD103 - non-T RM (H32-22) clones. Mean immunofluorescence intensity (MFI) are in parentheses. (B) Cytotoxic activity of the T RM clone (Heu171) towards autologous IGR-Heu, Heu-CSC, CSC-1 and CSC-2 target cells. Percent of specific lysis are shown at indicated effector to target (E:T) ratios. (C) Quantification of transforming growth factor (TGF)-beta in conditioned media from IGR-Heu, Heu-CSC, CSC-1 and CSC-2 by multi-analyte flow assay. Ratios of active/total TGF-beta normalized to IGR-Heu are included. Results are presented as mean+-SEM of duplicates. (D) Cytotoxicity of the non-T RM clone (H32-22) towards autologous IGR-Heu, Heu-CSC, CSC-1 and CSC-2 target cells. (E) Inhibition of T RM -cell-mediated killing. CSC-1 cells are preincubated in the presence of isotype control, anti-major histocompatibility complex class I (MHC-I) or anti-intercellular adhesion molecule 1 (ICAM-1) neutralizing monoclonal antibody (mAb) and then CTL were added at 5:1 E:T ratio. Symbols represent replicates and horizontal bars represent means+-SEM (n=3). P value was determined by two-way analysis of variance test. *p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Effects of TNF on the suppressive function of TNFR2 + Tregs. Purified CD4 + T cells were cultured in medium alone, TNF (50 ng/mL), TNF combined with anti-TNFR2 mAbs (10 ug/mL), or isotype IgG, as indicated, for 72 hours. The representative FACS analysis of (A) CTLA-4 + cells and (B) PD-L1 + cells in TNFR2 + Tregs as indicated, gated on live CD4 + Foxp3 + cells. Summary of the proportions of (C) CTLA-4 + cells (n=3) and (D) PD-L1 + cells (n=3) in Tregs within each condition. Data are expressed as means +- SEM. *, P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 T FR expansion in lymphoid tissues during chronic SIV infection. ( a ) Disaggregated lymph node and spleen cells from SIV uninfected ( n =9) or chronically SIV-infected rhesus macaques ( n =11) were analysed by flow cytometry. Representative examples of flow cytometry gating are shown. Of viable CD3 + CD8 - cells, follicular subsets were defined as CXCR5 + cells (F) and germinal centre subsets were defined as CXCR5 hi PD-1 hi cells (GC). Of these subsets, regulatory cells were defined as CD25 hi CD127 - . T FR (CXCR5 + CD25 hi CD127 - ) were Foxp3 + , whereas T FH (CXCR5 + CD25 lo/- ) were Foxp3 - . ( b ) The percentages of each rhesus macaque regulatory subset, as analysed in a are shown. ( c ) The ratios of each regulatory cell population to its non-regulatory cell counterpart are shown. ( d ) The percentage of total CTLA-4 expression is shown in SIV-uninfected ( n =9) and chronically SIV-infected ( n =8) rhesus macaques. The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Mann-Whitney (Wilcoxon) tests to compare unpaired, nonparametric values and significance is denoted by asterisks where * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 Flow cytometry analysis of T cell-/lymphocyte-specific markers on normal and malignant B cells from CLL patients. a Summary scheme representing functional implications of CLL-specific candidate genes selected for flow cytometric analysis. b Flow cytometric analysis of expression of CTLA-4, TIGIT, CD276, LILRB4, and CD2 on peripheral blood B cells of CLL patients. The expression was determined for non-malignant B cells (""Normal""; CD19 + CD5 - B cells, represented in green) and neoplastic B cells (""CLL"", CD19 + CD5 + B cells, represented in orange) detected in the same samples. ""Co,"" no antibody staining control; ""Ab,"" staining with the antibody of interest as indicated. c Normalized median fluorescence intensities (target MFI - MFI of negative control [Co]; nMFI). d Delta normalized median fluorescence intensities between CLL cells and normal B cells (DeltanMFI (CLL-normal)) for each patient tested

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 The intracellular AND logic with different signaling domains. a Diagram of intracellular AND logic. b Primary human CD8+ T cells were transduced with FOS zipCAR-containing CD3zeta domain and RR zipCAR-containing CD28 domain. Cytotoxicity against Her2- and Axl-expressing Nalm6 was measured 24 h after adding alpha-Her2-SYN9 and/or alpha-Axl-EE zipFvs. The heatmap indicates cytotoxicity at varying zipFv concentrations ( n = 3, data are represented as mean). c Cytotoxicity of CD8+ T cells transduced with FOS zipCAR-containing CD3zeta domain and RR zipCAR-containing 4-1BB domain. The heatmap indicates cytotoxicity at varying zipFv concentrations ( n = 3, data are represented as mean). d (Left) Isolated Treg cells were transduced with two zipCAR constructs: SYN6-CD3zeta-P2A-FOXP3 and SYN1-CD28-P2A-puro. After puromycin selection (2 mug/mL), Treg cells were co-cultured with Her2- and Axl-expressing Nalm6 target cells (Right) The heatmap shows surface CTLA-4 expression detected after 48 h by flow cytometry at varying zipFv concentrations (alpha-Axl-SYN5 and alpha-Her2-SYN2) ( n = 3, data are represented as mean).