CD1d Antibody | WTH1

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CD1d Antibody | WTH1 gallery image 1

Immunoperoxidase staining of LEW rat spleen follicle with Mouse anti Rat CD1d (MCA5799)

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CD1d Antibody | WTH1 gallery image 2

Staining of rat thymocytes with Mouse anti Rat CD1d:FITC (MCA5799F)

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CD1d Antibody | WTH1 gallery image 3

Published investigator image:
Mouse anti Rat CD1d antibody, clone WTH1 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.

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CD1d Antibody | WTH1 gallery image 4

Published investigator image:
Mouse anti Rat CD1d antibody, clone WTH1 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.

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CD1d Antibody | WTH1 gallery image 5

Published investigator image:
Mouse anti Rat CD1d antibody, clone WTH1 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.

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CD1d Antibody | WTH1 gallery image 6

Published investigator image:
Mouse anti Rat CD1d antibody, clone WTH1 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.

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CD1d Antibody | WTH1 gallery image 7

Published investigator image:
Mouse anti Rat CD1d antibody, clone WTH1 used to block NKT cell responsed 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.

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CD1d Antibody | WTH1 gallery image 8

Published investigator image:
Epitope mapping of Mouse anti Rat CD1d antibody, clone WTH1 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 aα1 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.

Enlarge
CD1d Antibody | WTH1 gallery image 9

Published investigator image:
Mouse anti Rat CD1d antibody 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.

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  • Mouse anti Rat CD1d
  • Mouse anti Rat CD1d:FITC
  • Mouse anti Rat CD1d
  • Mouse anti Rat CD1d:Biotin
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    WTH1
  • Isotype
    IgG2a
3 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA5799FFdatasheet pdfdatasheet pdf0.1 mg
    MCA5799F
    MCA5799GAC, E, F, FN*, IP, WB*datasheet pdfdatasheet pdf0.1 mg
    MCA5799GA
    MCA5799BC, E, F, FN*, IP, WB*datasheet pdfdatasheet pdf0.1 mg
    MCA5799B
    MCA5799C, E, F, FN*, IP, WB*datasheet pdfdatasheet pdf0.25 mg
    MCA5799
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • 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 recognizes a different, non-overlapping epitope to clone WTH2 and 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.
    • Intended Use
    • Target Species
      Rat
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Mouseyes
      Cotton ratno
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Purified IgG - liquid
      Purified IgM conjugated to Fluorescein Isothiocyanate Isomer 1 (FITC) - liquid
      Purified IgG - liquid
      Purified IgG conjugated to Biotin - liquid
    • Reconstitution
    • Preparation
      Pack Size: 0.1 mgPurified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
      Pack Size: 0.25 mgPurified IgG prepared by affinity chromatography on Protein A
      Purified IgG prepared by affinity chromatography on Protein A
      Pack Size: 0.1 mgPurified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
      Pack Size: 0.25 mgPurified IgG prepared by affinity chromatography on Protein A
      Purified IgG prepared by affinity chromatography on Protein A from tissue culture supernatant
    • Preservative Stabilisers
      0.02% Sodium Azide (NaN3)
      0.09% Sodium Azide (NaN3)
      0.02% Sodium Azide (NaN3)
      0.02% Sodium Azide (NaN3)
    • Immunogen
      Rat CD1d-transduced M12.4.1.C3 cells.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 0.5mg/ml
      IgG concentration 0.5mg/ml
      IgG concentration 0.5mg/ml
      IgG concentration 0.5mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
      Phosphate buffered saline
      Phosphate buffered saline
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. This product is photosensitive and should be protected from light.
      Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
      18 months from date of despatch.
      18 months from date of despatch.
      18 months from date of despatch.
    • GO Terms
      late endosome
      positive regulation of NK T cell activation
      endogenous lipid antigen binding
      positive regulation of interleukin-4 biosynthetic process
      positive regulation of interferon-gamma biosynthetic process
      integral to membrane
      protein binding
      positive regulation of interleukin-2 biosynthetic process
      positive thymic T cell selection
      antigen processing and presentation, exogenous lipid antigen via MHC class Ib
      positive regulation of NK T cell differentiation
      lysosomal membrane
      innate immune response
      MHC class I protein complex
      early endosome
      positive regulation of T cell mediated cytotoxicity
      external side of plasma membrane
      endosome membrane
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Flow Cytometry1/2,000
      Immunohistology - Frozen
      Immunoprecipitation
      Functional Assays(1)
      Western Blotting(2)
      (1)
      This antibody contains sodium azide, removal by dialysis is recommended prior to use in functional assays. Bio-Rad recommend the use of EQU003 for this purpose.
      (2)
      Clone WTH1 recognizes CD1d under non-reducing conditions, see Monzon-Casanova, E. et al. for details.
    • Application NameYesNoMin DilutionMax Dilution
      Flow Cytometry1/1,000
    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Flow Cytometry1/2,000
      Immunohistology - Frozen
      Immunoprecipitation
      Functional Assays(1)
      Western Blotting(2)
      (1)
      This antibody contains sodium azide, removal by dialysis is recommended prior to use in functional assays. Bio-Rad recommend the use of EQU003 for this purpose.
      (2)
      Clone WTH1 recognizes CD1d under non-reducing conditions, see Monzon-Casanova, E. et al. for details.
    • Application NameYesNoMin DilutionMax Dilution
      ELISA
      Flow Cytometry1/1,000
      Immunohistology - Frozen
      Immunoprecipitation
      Functional Assays(1)
      Western Blotting(2)
      (1)
      This antibody contains sodium azide, removal by dialysis is recommended prior to use in functional assays. Bio-Rad recommend the use of EQU003 for this purpose.
      (2)
      Clone WTH1 recognizes CD1d under non-reducing conditions, see Monzon-Casanova, E. et al. for details.

    • 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.
    • Technical Advice
    • Technical Advice
    • Technical Advice
    • Technical Advice
    • Recommended Protocol
    • Recommended Protocol
    • Recommended Protocol
    • Recommended Protocol
    • Flow Cytometry
      Use 10ul of the suggested working dilution to label 1x106 cells in 100ul.
    • ELISA
    • ELISA
    • ELISA
    • ELISA
    • Immunohistology
    • Immunohistology
    • Immunohistology
    • Immunohistology
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Immunofluorescence
    • Western Blotting
    • Western Blotting
    • Western Blotting
    • Western Blotting
    • Instructions For Use
    • Instructions For Use
    • Instructions For Use
    • Instructions For Use

    Additional CD1d Antibody Formats

    Formats Clone Applications Sizes available
    CD1d Antibody : Biotin WTH1 C, E, F, FN*, IP, WB* 0.1 mg
    CD1d Antibody : Purified WTH1 C, E, F, FN*, IP, WB* 0.1 mg | 0.25 mg
    CD1d Antibody : FITC WTH1 F 0.1 mg
    • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

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              References

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