GITRL antibody | YGL386
Expression of heterologous membrane proteins in BV fraction confirmed by Western blot. Ten micrograms of each of the BV samples expressing CD2 or its ligand CD58 (A), CD40 or CD40L (B), and GITR or GITRL (C) were loaded in each lane. The blotted membranes were immuno-stained with either anti-FLAG or anti-HA antibodies according to the attached tag as described in Materials and Methods. The positions of the molecular mass marker proteins are indicated on the left.
From: Sakihama T, Sato T, Iwanari H, Kitamura T, Sakaguchi S, et al. (2008) A Simple Detection Method for Low-Affinity Membrane Protein Interactions by Baculoviral Display. PLoS ONE 3(12): e4024.
Detection of specific interaction between CD40 and CD40L or GITR and GITRL displayed on BV. (A) Left panel: binding of CD40L-BV to the immobilized CD40-BV. Wells were coated with 0.25 µg/well of CD40-BV or wild-type BV. Binding was detected by using a biotinylated anti-mouse CD40L monoclonal antibody (clone MR1) and HRP-streptavidin. Right panel: binding of CD40-BV to the immobilized CD40L-BV. Wells were coated with 1 µg/well of CD40L-BV or wild-type BV. Binding was detected by using an anti-mouse CD40 monoclonal antibody (clone 3/23) and HRP-anti rat IgG+IgM. (B) Left panel: binding of GITRL-BV to the immobilized GITR-BV. Wells were coated with 1 µg/well of GITR-BV or wild-type BV. Binding was detected by using an anti-mouse GITRL monoclonal antibody (clone YGL386) and HRP-anti rat IgG+IgM. Right panel: binding of GITR-BV to the immobilized GITRL-BV. Wells were coated with 1 μg/well of GITRL-BV or wild-type BV. Binding was detected by using an anti-mouse GITR monoclonal antibody (clone DTA-1) and HRP-anti rat IgG+IgM. Filled circles indicate binding to BV displaying respective receptors (or ligands). Open circles indicate binding to wild-type BV.
From: Sakihama T, Sato T, Iwanari H, Kitamura T, Sakaguchi S, et al. (2008)
A Simple Detection Method for Low-Affinity Membrane Protein Interactions by Baculoviral Display.
PLoS ONE 3(12): e4024.
Specific binding of ligand-displaying BV to cells expressing receptor. (A) Schematic view. X corresponds to CD2 (for B), CD40 (for C), or GITR (for D). Y corresponds to CD58 (for B), CD40L (for C), or GITRL (for D). (B) CD58-displaying BV binds to CD2-positive (top), but not to CD2-negative Jurkat cells (bottom). Left: binding of an anti-CD2 monoclonal antibody (clone RPA-2.10) plus a PE-anti mouse Ig-κ chain antibody. Right: binding of CD58-displaying BV plus an anti-gp64 antibody (clone A0505A) and a PE-anti mouse Ig-κ chain antibody. The thin line indicates cells incubated with the 2nd antibody only. (C) CD40-dependent binding of CD40L-displaying BV to mouse splenic B cells. BALB/c mouse splenocytes were incubated with CD40L-displaying BV and a biotinylated anti-gp64 monoclonal antibody (clone B8147A), and then were stained with FITC-labeled streptavidin and a PE-labeled anti-B220 antibody. CD40L-BV binding to the B cells (shown in the center lower panel) was blocked by pre-incubation with an anti-mouse CD40 monoclonal antibody (clone HM40-3) (the right upper panel) but not by control hamster IgG (right lower panel). (D) Binding of GITRL-displaying BV to GITR-positive cells. GITR-expressing mouse T cell hybridoma 18.3.5 cells were incubated with GITRL-displaying BV (right) or wild-type BV (left). Binding of BV was detected with a biotinylated anti-gp64 monoclonal antibody B8147A and PE-streptavidin. The thin line indicates cells incubated without BV.
From: Sakihama T, Sato T, Iwanari H, Kitamura T, Sakaguchi S, et al. (2008)
A Simple Detection Method for Low-Affinity Membrane Protein Interactions by Baculoviral Display.
PLoS ONE 3(12): e4024.
Rat anti Mouse GITRL antibody, clone YGL386 recognizes mouse glucocorticoid-induced tumour necrosis factor receptor ligand (GITRL), a 20 kDa member of the tumour necrosis factor ligand superfamily, otherwise known as TNSF18.
Mouse GITRL is predominantly expressed on immature and mature dendritic cells, macrophages and B cells. Expression of mouse GITRL can be transiently up regulated on these cells, following lipolysaccharide (LPS) stimulation, but protein expression is rapidly down modulated shortly after stimulation.
Studies demonstrate that GITRL can neutralise the suppressive effects of CD4+CD25+ regulatory T cells and can also provide a co-stimulatory signal for both naïve and primed T cells, suggesting a key role for GITRL in the regulation of T cell-mediated immunity.
Mouse GITRL is predominantly expressed on immature and mature dendritic cells, macrophages and B cells. Expression of mouse GITRL can be transiently up regulated on these cells, following lipolysaccharide (LPS) stimulation, but protein expression is rapidly down modulated shortly after stimulation.
Studies demonstrate that GITRL can neutralise the suppressive effects of CD4+CD25+ regulatory T cells and can also provide a co-stimulatory signal for both naïve and primed T cells, suggesting a key role for GITRL in the regulation of T cell-mediated immunity.
Product Details
- Target Species
- Mouse
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
0.09% sodium azide - Carrier Free
- Yes
- Immunogen
- Recombinant mouse GITRL.
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised DA rats were fused with cells of the Y3/Ag1.2.3 myeloma cell line.
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Guarantee
- 18 months from date of despatch.
More Information
- UniProt
- Q9UNG2 Related reagents
- Entrez Gene
- TNFSF18 Related reagents
- GO Terms
- GO:0016021 integral to membrane
- GO:0007165 signal transduction
- GO:0005125 cytokine activity
- GO:0005164 tumor necrosis factor receptor binding
- GO:0005615 extracellular space
- GO:0006916 anti-apoptosis
- GO:0006955 immune response
- GO:0007267 cell-cell signaling
- Regulatory
- For research purposes only
Applications of GITRL antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA | |||
| Flow Cytometry | 1/25 | 1/50 |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 106 cells in 100ul.
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| Goat anti Rat IgG:HRP (Mouse Adsorbed) | STAR72 | C E P | 0.5 mg |
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| Goat F(ab')2 anti Rat IgG:RPE (Mouse Adsorbed) | STAR73 | F | 0.5 ml |
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Product Specific References
References for GITRL antibody
-
Cobbold, S.P. et al. (2004) Induction of foxP3+ regulatory T cells in the periphery of T cell receptor transgenic mice tolerized to transplants.
J Immunol. 172 (10): 6003-10. -
Sakihama, T. et al. (2008) A simple detection method for low-affinity membrane protein interactions by baculoviral display.
PLoS One. 3 (12): e4024. -
Tone, M. et al. (2003) Mouse glucocorticoid-induced tumor necrosis factor receptor ligand is costimulatory for T cells.
Proc Natl Acad Sci U S A. 100 (25): 15059-64.
