CD1d antibody | WTH1
Mouse anti Rat CD1d antibody, clone WTH1 (MCA5799) used for the evaluation of CD1d expression on rat cell targets by flow cytometry
Image caption:
CD1d cell surface expression in five different rat strains. CD1d cell surface expression was analyzed by flow cytometry using the biotinylated WTH-1 mAb visualized with SA-APCy in thymocytes and splenocytes derived from F344, PVG, BN, LEW and DA rats.
From: Monzon-Casanova E, Steiniger B, Schweigle S, Clemen H, Zdzieblo D, Starick L, et al. (2010)
CD1d Expression in Paneth Cells and Rat Exocrine Pancreas Revealed by Novel Monoclonal Antibodies Which Differentially Affect NKT Cell Activation.
PLoS ONE 5(9): e13089.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Rat CD1d antibody, clone WTH1 (MCA5799) used for the evaluation of CD1d expression on rat and mouse cell targets by flow cytometry, immunoprecipitation and western blotting.
Image caption:
Characterization of two novel anti-CD1d monoclonal antibodies.
(A) Titration of WTH-1 and WTH-2 mAbs on rat CD1d or mouse CD1d1 Raji transductants (left) and on C57BL/6 or LEW thymocytes (right). Cells were stained with the indicated concentrations of CD1d-specific antibodies (X axis). After washing, antibodies were detected with PE-labeled donkey anti-mouse IgG and analyzed by flow cytometry. On the Y axis, the geometric mean fluorescence intensity (MFI) is shown for the different antibody concentrations. (B) Immunoprecipitation of biotinylated surface proteins with WTH-1 (1), WTH-2 (2) or isotype control antibodies. Immunoprecipitated material was size separated under reducing conditions on a 15% SDS-PAGE, blotted onto a membrane and detected by addition of streptavidin-HRP. (C) Western blot analysis of proteins derived from rat CD1d (r) or mouse CD1d1 (m) transduced cells - left blots - and from rat tissues: spleen (spl), thymus (thy) and pancreas (pan) - right blots -. Proteins were separated on a 10% polyacrylamide gel under non-reducing conditions and blotted to a membrane. CD1d was detected with the WTH-1 or WTH-2 mAbs. The duration of film exposure for the CD1d immunoblots is indicated under the mAb names. After CD1d detection, the blots were stripped and re-probed with a polyclonal ERK2-specific antibody as protein loading control (lower blots).
From: Monzon-Casanova E, Steiniger B, Schweigle S, Clemen H, Zdzieblo D, Starick L, et al. (2010)
CD1d Expression in Paneth Cells and Rat Exocrine Pancreas Revealed by Novel Monoclonal Antibodies Which Differentially Affect NKT Cell Activation.
PLoS ONE 5(9): e13089.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Rat CD1d antibody, clone WTH1 (MCA5799) used for the evaluation of CD1d expression on rat cell targets by immunohistochemistry on cryostat sections.
Image caption:
CD1d distribution in lymphatic organs.
ABC technique with unconjugated mAb WTH-1 unless otherwise stated. (A) LEW thymus. Bar=500 μm. (B) Longitudinal section of LEW spleen. Hemalum counterstain. Bar=250 μm. (C) Cross-section of LEW splenic white pulp. WTH-1 biotin-conjugated primary antibody. Bar=250 μm. (D) C57BL/6 mouse thymus. Arrows indicate cortico-medullary boundary. Biotin-conjugated antibody. Bar=250 μm. (E) HIS57 staining in LEW splenic white pulp. Bar=250 μm. (F) CD4 (mAb W3/25) expression in LEW spleen. Bar=500 μm. Abbreviations in lymphatic organs: m, medulla; c, cortex; rp, red pulp; pals, periarteriolar lymphatic sheath; mz, marginal zone and f, follicle.
From: Monzon-Casanova E, Steiniger B, Schweigle S, Clemen H, Zdzieblo D, Starick L, et al. (2010)
CD1d Expression in Paneth Cells and Rat Exocrine Pancreas Revealed by Novel Monoclonal Antibodies Which Differentially Affect NKT Cell Activation.
PLoS ONE 5(9): e13089.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Rat CD1d antibody, clone WTH1 (MCA5799) used to block NKT cell responses in vitro
Image caption:
Figure 5. Effects of anti-CD1d mAbs on antigen presentation to type I and II NKT cells.
(A) Blocking of type I NKT cells responses by the mAbs WH1-1 and WTH-2. IL-4 production was analyzed by measuring cytokine release into the culture supernatant 24 hours after initiation of the cultures by ELISA. Total splenocytes from C57BL/6 mice or F344 rats were cultured with α-GalCer (10 ng/ml), in the presence or absence of WTH-1, WTH-2 or isotype control antibodies (with a final saturating concentration of 3.6 μg/ml). Results were normalized to 100% for IL-4 production without antibody. Bars show means + standard deviation (SD) of the results obtained in three independent experiments. IL-4 concentrations without antibody were 9.9, 3.3 and 6.5 pg/ml and 15.8, 28.4 and 20.9 pg/ml for rat and mouse cultures, respectively. (B) Effects of the antibodies on CD1d antigen presentation to type II NKT cells. IL-2 was measured by ELISA in supernatants of co-cultures of type II NKT cell hybridomas and CD1d-transduced cells. Raw data were normalized to IL-2 production without antibody equaling 100%. The graphics represent the mean values + SD obtained from three independent experiments. The left graph shows IL-2 production by 5×104 VIII24.1 T cell hybridoma cells after co-culture with 5×104 mCD1d1-transduced LBB cells. IL-2 concentrations without addition of antibodies, in the three independent experiments, were 575, 415 and 572 pg/ml, respectively. The right graph gives the corresponding results for 1×104 XV19.2 T cell hybridoma cells after co-culture with 5×104 mCD1d1 transduced Raji cells. IL-2 concentrations without added antibody in the three independent experiments were 33.9, 101.6 and 22.6 pg/ml, respectively. To determine differences between cultures where isotype control antibody or anti-CD1d mAbs were added, two-tailed paired t-test was used. * and ** indicate p-values <0.05 and <0.005, respectively.
From: Monzon-Casanova E, Steiniger B, Schweigle S, Clemen H, Zdzieblo D, Starick L, et al. (2010)
CD1d Expression in Paneth Cells and Rat Exocrine Pancreas Revealed by Novel Monoclonal Antibodies Which Differentially Affect NKT Cell Activation.
PLoS ONE 5(9): e13089.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Epitope mapping of Mouse anti Rat CD1d antibody, clone WTH1(MCA5799) by flow cytometry
Image caption:
Epitope mapping of CD1d-specific mAbs.(A) Alignment of the amino acid sequences of the extracellular domains of the CD1d molecules used in this study. α-helical regions are illustrated in bold. The open triangle points out the localization of the rat CD1d allelism. Mutations in mouse CD1d1 are highlighted with closed triangles. The arrow indicates where mouse/human chimeras were joined. Numbers show rat CD1d amino acid positions. Accession numbers for the amino acid sequences can be found in GenBank: rCD1d (Rattus norvegicus CD1d), BAA82323; mCD1d1 (Mus musculus CD1d1), NP_031665; sCD1d1 (Mus spretus CD1d1), ACM45455; hCD1d1 (Homo sapiens CD1d), NP_001757. The mCD1d2 (Mus musculus CD1d2) amino acidic sequence differs in one amino acid in the signal peptide (tryptophan, position −13) from the sequence published under the accession number: P11610. (B) Binding capacity of mAbs to different CD1d molecules. Raji cells were transduced with CD1d molecules using a retroviral expression system. Bicistronic EGFP in the CD1d expression vectors served as reporter gene. Cells were stained with unconjugated WTH-1 or WTH-2 mAbs followed by PE-labeled donkey anti-mouse IgG or PE-labeled 1B1 mAb and were analyzed by flow cytometry. All primary antibodies were used at a final concentration of 250 ng/ml. (C) Localization of relevant amino acids in the CD1d tertiary structure. The model depicts part of the co-crystal of the PBS25 glycolipid and mouse CD1d (PDB: 3GMP) and it was visualized using PDB Swiss Viewer Deep View v4.0 [52] (http://www.expasy.org/spdbv/). The αα1 and α2 domains are shown. Gray and green ribbon diagrams highlight the regions constituting the N- and C-terminal parts, respectively, of the chimeric molecules. Spheres visualize the amino acids discussed in the text: aspartic acid (D) 93 in red, methionine (M) 162 in green, valine (V) 118 in gray and arginine (R) 21 in blue.
From: Monzon-Casanova E, Steiniger B, Schweigle S, Clemen H, Zdzieblo D, Starick L, et al. (2010)
CD1d Expression in Paneth Cells and Rat Exocrine Pancreas Revealed by Novel Monoclonal Antibodies Which Differentially Affect NKT Cell Activation.
PLoS ONE 5(9): e13089.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Rat CD1d antibody, clone WTH1 (MCA5799) used to evaluate CD1d expression on mouse, rat and cotton rat cells by flow cytometry. Results indicate the lack of cross-reactivity of the WTH-1 clone with the cotton rat (Sigmodon hispidus).
Image caption:
WTH‐2 and 1B1 but not WTH‐1 monoclonal CD1d antibodies can be used to stain cotton rat CD1d. Representative data from one of two experiments in total are shown. The biotinylated monoclonal mouse anti‐rat antibodies WTH‐1 and WTH‐2 (each 125 ng/ml), detected with streptavidin‐allophycocyanin, and rat anti‐mouse 1B1 phycoerythrin (2 μg/ml) were used to stain Raji cells transduced with rat, mouse, or cotton rat CD1d. Staining intensities are compared to isotype controls.
From: Fichtner AS, Paletta D, Starick L, Schumann RF, Niewiesk S, Herrmann T.
Function and expression of CD1d and invariant natural killer T-cell receptor in the cotton rat (Sigmodon hispidus).
Immunology. 2015 Dec;146(4):618-29.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Rat CD1d antibody, clone WTH1 (MCA5799) used for the evaluation of CD1d expression on rat cell targets by immunohistochemistry on cryostat sections.
Image caption:
CD1d distribution in non-lymphatic organs.
ABC technique with unconjugated mAb WTH-1 unless otherwise stated.(G) LEW liver. Bar=250 μm. (H) LEW heart. Hemalum counterstain. Bar=250 μm. (I) LEW ileum. The epithelium at the tips of the villi (left rim of picture) is not preserved. Biotinylated WTH-1 and hemalum counterstain. Bar=100 μm. (J) C57BL/6 ileum. Arrows indicate CD1d positive enteroendocrine cells in the epithelium. Biotin-conjugated primary antibody, tyramide-amplified ABC technique and hemalum counterstain. Bar=50 μm. (K) C57BL/6 CD1d−/− ileum. Biotin-conjugated primary antibody, tyramide-amplified ABC technique and hemalum counterstain. Bar=50 μm. (L) LEW pancreas. Biotinylated WTH-1. Bar=50 μm. (M) C57BL/6 pancreas. Biotin-conjugated primary antibody, tyramide-amplified ABC technique. and hemalum counterstain. Bar=50 μm. (N) C57BL/6 CD1d−/− pancreas. Biotin-conjugated primary antibody, tyramide-amplified ABC technique and hemalum counterstain. Bar=50 μm. Abbreviations in non-lymphatic organs: c, crypts; lp, lamina propria; m, smooth muscle cells of the gut wall; i, islet of Langerhans and d, interlobular duct.
From: Monzon-Casanova E, Steiniger B, Schweigle S, Clemen H, Zdzieblo D, Starick L, et al. (2010)
CD1d Expression in Paneth Cells and Rat Exocrine Pancreas Revealed by Novel Monoclonal Antibodies Which Differentially Affect NKT Cell Activation.
PLoS ONE 5(9): e13089.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Mouse anti Rat CD1d:Biotin
Mouse anti Rat CD1d:FITC
Mouse anti Rat CD1d
- Product Type
- Monoclonal Antibody
- Clone
- WTH1
- Isotype
- IgG2a
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| MCA5799B | C E F IP WB* | 0.1 mg |
|
|
| MCA5799F | F | 0.1 mg |
|
|
| MCA5799GA | C E F FN* IP WB* | 0.1 mg |
|
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| MCA5799 | C E F FN* IP WB* | 0.25 mg |
|
Mouse anti Rat CD1d antibody, clone WTH1 recognizes rat CD1d, a glycoprotein expressed on cortical thymocytes, certain T-cell leukaemias and various other antigen presenting cells. CD1d binds self and non-self glycolipids and presents them to T-cell receptors on natural killer cells. When activated, natural killer cells produce Th1 and Th2 cytokines.
Clone WTH1 binds with high affinity to the mature CD1d heavy chain independently of beta-2-microglobulin. Mouse anti Rat CD1d antibody, clone WTH1 efficiently blocks rat and mouse invariant natural killer T cell (iNKT cell) response to a-GalCer (an antigen for iNKT cells) and has a slightly higher affinity to native CD1d than clone WTH2.
Clone WTH1 cross reacts weakly with mouse tissue in IHC and western blotting, but reacts strongly with both rat and mouse antigens in flow cytometry and immunoprecipitation.
Clone WTH1 binds with high affinity to the mature CD1d heavy chain independently of beta-2-microglobulin. Mouse anti Rat CD1d antibody, clone WTH1 efficiently blocks rat and mouse invariant natural killer T cell (iNKT cell) response to a-GalCer (an antigen for iNKT cells) and has a slightly higher affinity to native CD1d than clone WTH2.
Clone WTH1 cross reacts weakly with mouse tissue in IHC and western blotting, but reacts strongly with both rat and mouse antigens in flow cytometry and immunoprecipitation.
Product Details
- Target Species
- Rat
- Species Cross-Reactivity
-
Target Species Cross Reactivity Mouse Cotton rat - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG conjugated to Biotin - liquid
- Product Form
- Purified IgM conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Buffer Solution
- Phosphate buffered saline
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- <0.1% Sodium Azide (NaN3)
- Preservative Stabilisers
- <0.1% Sodium Azide (NaN3)
- Preservative Stabilisers
- <0.1% Sodium Azide (NaN3)
- Immunogen
- Rat CD1d-transduced M12.4.1.C3 cells.
- Approx. Protein Concentrations
- IgG concentration 0.5mg/ml
- Approx. Protein Concentrations
- IgG concentration 0.5mg/ml
- Approx. Protein Concentrations
- IgG concentration 0.5mg/ml
Storage Information
- Storage
- This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. - Storage
- This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. This product is photosensitive and should be protected from light. - Storage
- This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended. - Guarantee
- 12 months from date of despatch
- Guarantee
- 12 months from date of despatch
- Guarantee
- 12 months from date of despatch
More Information
- UniProt
- P11609
- Q63493
- Entrez Gene
- Cd1d1
- Cd1d1
- GO Terms
- GO:0005515 protein binding
- GO:0001916 positive regulation of T cell mediated cytotoxicity
- GO:0016021 integral to membrane
- GO:0005765 lysosomal membrane
- GO:0005769 early endosome
- GO:0005770 late endosome
- GO:0009897 external side of plasma membrane
- GO:0010008 endosome membrane
- GO:0030883 endogenous lipid antigen binding
- GO:0045059 positive thymic T cell selection
- GO:0045078 positive regulation of interferon-gamma biosynthetic process
- GO:0045086 positive regulation of interleukin-2 biosynthetic process
- GO:0045087 innate immune response
- GO:0045404 positive regulation of interleukin-4 biosynthetic process
- GO:0048007 antigen processing and presentation, exogenous lipid antigen via MHC class Ib
- GO:0051135 positive regulation of NK T cell activation
- GO:0051138 positive regulation of NK T cell differentiation
- GO:0042612 MHC class I protein complex
- Regulatory
- For research purposes only
Applications of CD1d antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA | |||
| Flow Cytometry | 1/1,000 | ||
| Immunohistology - Frozen | |||
| Immunoprecipitation | |||
| Western Blotting 1 | |||
| Flow Cytometry | 1/1,000 | ||
| ELISA | |||
| Flow Cytometry | 1/2,000 | ||
| Functional Assays 1 | |||
| Immunohistology - Frozen | |||
| Immunoprecipitation | |||
| Western Blotting 2 |
- 1Clone WTH1 recognizes CD1d under non-reducing conditions, see Monzon-Casanova, E. et al. for details.
- 1 This antibody contains sodium azide, removal by dialysis is recommended prior to use in functional assays.
- 2 Clone WTH1 recognizes CD1d under non-reducing conditions (Monzon-Casanova, E. et al.).
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.
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.
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 1x106 cells in 100ul.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
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Secondary Antibodies Available
Negative Isotype Controls Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Mouse IgG2a Negative Control:FITC | MCA1210F | F | 0.1 mg |
|
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| Mouse IgG2a Negative Control | MCA1210 | F | 100 Tests |
|
Product Specific References
References for CD1d antibody
-
Monzon-Casanova, E. et al. (2010) CD1d expression in paneth cells and rat exocrine pancreas revealed by novel monoclonal antibodies which differentially affect NKT cell activation.
PLoS One 5:e13089 -
Fichtner AS et al. (2015) Function and expression of CD1d and invariant natural killer T-cell receptor in the cotton rat (Sigmodon hispidus).
Immunology. 146 (4): 618-29. -
Monzon-Casanova, E. et al. (2013) Direct identification of rat iNKT cells reveals remarkable similarities to human iNKT cells and a profound deficiency in LEW rats.
Eur J Immunol. 43 (2): 404-15.
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