CTLA4
Reactivity: Human, Mouse, Rat
FACS, WB, ELISA, IHC (p), IF (p)
Host: Rabbit
Polyclonal
unconjugated
Application Notes
For flow cytometric applications, a three step labeling procedure is recommended for amplifying signal. Suggested protocol for 3-step staining using concanavalin-A-stimulated peripheral blood mononuclear cells method: 1. Incubate 100 µl concanavalin-A-stimulated 3-day peripheral blood mononuclear cells (1x10^6) with primary (unconjugated) antibody for 20-30 minutes at room temperature. 2. Add 2 mls of 1X lysing buffer (10X Pharm Lyse™ Lysing Buffer, Cat. No. 555899) and incubate for 10-15 minutes. Centrifuge and aspirate. 3. Wash once with PBS/0.1% sodium azide/1% heat-activated fetal bovine serum (PBS-FBS). Centrifuge and aspirate. 4. Add biotinylated goat anti-mouse Ig (Cat. No. 553999) and incubate for 20-30 minutes at room temperature. 5. Wash once with PBS-FBS. Centrifuge and aspirate. 6. Add SAV-PE (Cat. No. 554061) and incubate for 20-30 minutes in the dark at room temperature. 7. Wash once with PBS-FBS. Centrifuge and aspirate. Resuspend in 0.5 ml of PBS-FBS and analyze by flow cytometry.
Restrictions
For Research Use only
Format
Liquid
Concentration
1.0 mg/mL
Buffer
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2μm sterile filtered.
Preservative
Azide free
Storage
4 °C
Storage Comment
Store undiluted at 4°C. This preparation contains no preservatives, thus it should be handled under aseptic conditions.
Morton, Fu, Stewart, Giacoletto, White, Leysath, Evans, Shieh, Karr: "Differential effects of CTLA-4 substitutions on the binding of human CD80 (B7-1) and CD86 (B7-2)." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 156, Issue 3, pp. 1047-54, (1996) (PubMed).
Kuiper, Brouwer, Linsley, van Lier: "Activated T cells can induce high levels of CTLA-4 expression on B cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 155, Issue 4, pp. 1776-83, (1995) (PubMed).
Lindsten, Lee, Harris, Petryniak, Craighead, Reynolds, Lombard, Freeman, Nadler, Gray: "Characterization of CTLA-4 structure and expression on human T cells." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 151, Issue 7, pp. 3489-99, (1993) (PubMed).
Target
CTLA4
(Cytotoxic T-Lymphocyte-Associated Protein 4 (CTLA4))
Reacts with the cytolytic T lymphocyte-associated antigen, CTLA-4. CTLA-4 is transiently expressed on activated CD28+ T cells and binds to CD80 and CD86 present on antigen presenting cells (APC) with high avidity. This interaction appears to deliver a negative regulatory signal to the T cell. There are recent reports that indicate that CTLA-4 is also expressed on B cells when cultured with activated T cells, suggesting a possible role of CTLA-4 in the regulation of B-cell response. Immobilized BNI3.1 enhances T-cell proliferation induced by CD3 and CD28. Recent studies have showed that CD152 can be expressed by regulatory T (Treg) cells. It has been found this antibody can stain the intracellular CD152 on the Treg cells after fixation and permeabilization of cells. Synonyms: CTLA-4