CD335 Antibody | AKS1

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CD335 Antibody | AKS1 gallery image 1

Staining of bovine peripheral blood lymphocytes with Mouse anti Bovine CD335 (MCA2365GA) followed by Goat anti Mouse IgG (H/L):FITC (STAR117F). A small positive peak representing NK cells can be identified

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CD335 Antibody | AKS1 gallery image 2

Published customer image:
Mouse anti bovine CD335 antibody, clone AKS1 used for the isolation by immunomagnetic separation and evaluation of CD335 expression by flow cytometry of NKp46 expression on bovine peripheral blood mononuclear cells.
Image caption:
CD2 distribution on bovine NK cells in peripheral blood and during cell culture. (A) CD2 expression by PBMCs from healthy cattle, and in NKp46-selected cells cultured for the indicated time in the presence of 100U/ml rbIL-2. Results were obtained by two-colour flow cytometry, gating for viable lymphocytes. Data shown are from one animal representative of 19. (B) Monitoring of cell divisions of CD2- and CD2+ NK cell populations during three and six days of rbIL-2 (100U/ml) stimulation. NKp46-selected cells were labelled with CFSE the day after isolation and CFSE intensity (filled histograms) measured in flow cytometry on the indicated days, gating for CD2- or CD2+ viable cells. Solid line open histograms show non-dividing NK cells, cultured in the presence of 1U/ml rbIL-2, while broken line open histograms show unlabeled stimulated NK cells. Data shown are from one animal representative of five. (C) Stability of CD2 expression in NK cell subsets cultured separately. NK cells separated with immunomagnetic beads for absence or presence of CD2 were cultured separately for eleven days in the presence of IL-2, monitoring CD2 expression at three different time points (filled histograms). Day 1 corresponds to the day of separation, 24 h after primary NK cell isolation. Open histograms show secondary mAb controls. Data shown are from one animal representative of eight.

From: Boysen P, Olsen I, Berg I, Kulberg S, Johansen GM, Storset AK. Bovine CD2-/NKp46+ cells are fully functional natural killer cells with a high activation status. BMC Immunol. 2006 Apr 27;7:10.

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CD335 Antibody | AKS1 gallery image 3

Published customer image:
Mouse anti Bovine CD335 antibody, clone AKS1 used as a positive antibody control in blocking experiments looking at lytic and γ interferon responses or bovine NK cells.
Image caption:
Effect of anti-CD2 mAbs on bovine NK cell functions. (A) IFN-γ response from NK cells caused by cross-linking of CD2. Plastic wells were pre-coated with two different mAbs against bovine CD2, along with NKp46-mAb as positive control and CD8-mAb or medium as negative controls. rbIL-2 activated NK cells were added and incubated for 24 h in the presence or absence of rbIL-12 (0.5 U/ml), and IFN-γ content in the supernatants analyzed by an ELISA. Data shown are means ± SD of two to four individual animals. (B-C) IL-2 activated NK cell killing of the bovine target lines BL3.1 and MDBK in a 4 h 51Cr-release assay, with addition of the indicated mAbs for blocking. CD2mAb I = BAQ95A, CD2 mAb II = MUC2A, CD8 mAb = CACT80C, NKp46 mAb = AKS1. (D) As above, using murine Fc-receptor bearing P815 target cells to obtain a redirected lysis effect. Data shown in (B-D) are from one animal representative of three to five.

From: Boysen P, Olsen I, Berg I, Kulberg S, Johansen GM, Storset AK. Bovine CD2-/NKp46+ cells are fully functional natural killer cells with a high activation status. BMC Immunol. 2006 Apr 27;7:10.

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CD335 Antibody | AKS1 gallery image 4

Published customer image:
Mouse anti bovine CD335 antibody used for the evaluation of NKp46 expression on bovine peripheral blood mononuclear cells by flow cytometry
Image caption:
Oenothein B Primes bovine CD335+ cells to respond to IL-18. (A) Bovine PBMCs (105 cells/well) were depleted of CD335+ cells and treated with 20 μg/ml oenothein B or X-VIVO medium alone for 24 hrs. Cells were then washed and treated with 10 ng/ml rhu IL-18 or medium alone for 18 hrs. After incubation, IFNγ levels in the supernatant fluids were measured by ELISA. The data are expressed as mean +/- SEM of three independent experiments comparing depleted PBMCs to un-depleted controls tested concurrently. All samples were tested in duplicate or triplicate. Statistical significance was measured by Two-way ANOVA with Bonferroni post-test. *p<0.05, **p<0.01, ***p<0.001 (B) Bovine PBMCs (105 cells/well) from a new calf were treated with the indicated amounts of oenothein B or X-VIVO medium alone for 24 hrs. Cells were washed and treated with 10 ng/ml rhu IL-18 or X-VIVO medium alone for 6 hrs in the presence of brefeldin A. IFNγ production was measured by intracellular flow cytometry. The data are expressed as mean+/- SEM. All samples were tested in triplicate. Statistical significance was measured by Two-way ANOVA with Bonferroni post-test. *p<0.05, **p<0.01, ***p<0.001 (C) Representative examples of two-color flow cytometry plots comparing IFNγ staining on CD335+ cells.

From: Ramstead AG, Schepetkin IA, Quinn MT, Jutila MA (2012) Oenothein B, a Cyclic Dimeric Ellagitannin Isolated from Epilobium angustifolium, Enhances IFN? Production by Lymphocytes.
PLoS ONE 7(11): e50546.

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CD335 Antibody | AKS1 gallery image 5

Published customer image:
Mouse anti bovine CD335 antibody used for the evaluation of NKp46 expression on bovine peripheral blood mononuclear cells by flow cytometry
Image caption:
Oenothein B induces IL-2Rα or CD69 on bovine and human NK cells. (A) Bovine PBMCs (105 cells/well) were treated with 20 μg/ml oenothein B in X-VIVO medium for 24 hrs, and IL-2Rα expression on NK cells was measured by multi-color flow cytometry. Representative examples of two-color flow cytometry plots comparing IL-2Rα staining on oenothein B-treated and untreated bovine NK cells (CD335+) from each animal are shown. (B) Human PBMCs (105 cells/well) were treated with 40 μg/ml oenothein B in cRPMI medium for 48 hrs. CD69 expression on NK cells was then measured by flow cytometry. Representative examples of two-color flow cytometry plots comparing CD69 staining on oenothein B-treated and untreated human NK cells from each donor are shown.

From: Ramstead AG, Schepetkin IA, Quinn MT, Jutila MA (2012) Oenothein B, a Cyclic Dimeric Ellagitannin Isolated from Epilobium angustifolium, Enhances IFNγ Production by Lymphocytes.
PLoS ONE 7(11): e50546.

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CD335 Antibody | AKS1 gallery image 6

Published customer image:
Mouse anti CD355 antibody, clone AKS1 used to evaluate NKp46 expression on bovine peripheral blood mononuclear cells by flow cytometry.
Image caption:
PBMC from a TB-reactor animal were stimulated with PPDB and flow cytometry analysis performed to enumerate IL-22 and IL-17A production in different lymphocyte populations. Numbers represent the percentage of single, live bovine lymphocytes. Data representative of at least 4 independent experiments.

From: Steinbach S, Vordermeier HM, Jones GJ.
CD4+ and ?δ T Cells are the main Producers of IL-22 and IL-17A in Lymphocytes from Mycobacterium bovis-infected Cattle.
Sci Rep. 2016 Jul 18;6:29990.

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  • Mouse anti Bovine CD335:RPE
  • Mouse anti Bovine CD335:Low Endotoxin
  • Mouse anti Bovine CD335:Alexa Fluor® 488
  • Mouse anti Bovine CD335
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    AKS1
  • Isotype
    IgG1
4 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    MCA2365GAF, IF, WBdatasheet pdfdatasheet pdf0.1 mg
    MCA2365GA
    MCA2365ELF, FN, IF, WBdatasheet pdfdatasheet pdf0.5 mg
    MCA2365EL
    MCA2365PEFdatasheet pdfdatasheet pdf100 Tests
    MCA2365PE
    MCA2365A488Fdatasheet pdfdatasheet pdf100 Tests/1ml
    MCA2365A488
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Mouse anti Bovine CD335 antibody, clone AKS1 recognizes bovine CD335, also known as NKp46 and Natural cytotoxicity triggering receptor 1. CD335 is a type I transmembrane protein, with two extracellular C2-type immunoglobulin-like domains, which functions as an activating receptor. CD335 is expressed by human natural killer cells (Sivori et al. 1997). The bovine homologue is expressed on bovine NK cells (Storset et al. 2004) and no expression of CD335 has been detected on B cells, T cells, monocytes or granulocytes.

      Clone AKS1 is reported to activate lysis of FcgammaR-expressing cell line P815, by IL-2 activated NKp46+ cells (Storset et al. 2004).
    • Intended Use
    • Target Species
      Bovine
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      American Bisonyes
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Purified IgG conjugated to R. Phycoerythrin (RPE) - lyophilised
      Purified IgG - liquid
      Purified IgG conjugated to Alexa Fluor® 488 - liquid
      Purified IgG - liquid
    • Reconstitution
      Reconstitute with 1 ml distilled water
    • Preparation
      Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
      Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
      Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
      Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
    • Preservative Stabilisers
      0.09%Sodium Azide
      1%Bovine Serum Albumin
      5%Sucrose
      None present
      0.09%Sodium Azide
      1%Bovine Serum Albumin
      0.09% Sodium Azide (NaN3)
    • Immunogen
      Fusion protein consisting of the extracellular region of bovine CD335.
    • Purity
    • Approx. Protein Concentrations
      IgG concentration 1.0 mg/ml
      IgG concentration 0.05 mg/ml
      IgG concentration 1.0 mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
      Phosphate buffered saline
      Phosphate buffered saline
      Phosphate buffered saline
    • Fusion Partners
      Spleen cells from immunised Balb/c mice were fused with cells of the NS-0 myeloma cell line.
    • Storage
      Prior to reconstitution store at +4oC.
      Following reconstitution store at +4oC.
      DO NOT FREEZE.
      This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use.
      Store at -20oC only.

      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.
      Store at +4oC or at -20oC if preferred.

      This product should be stored undiluted.

      Storage in frost-free freezers is not recommended. 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.

      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.
    • Shelf Life
      12 months from date of reconstitution.
      18 months from date of despatch.
      18 months from date of despatch.
      18 months from date of despatch.
    • GO Terms
      integral to membrane
      plasma membrane
      receptor activity
    • UniProt
    • Entrez Gene
    • Acknowledgements
      This product is provided under an intellectual property licence from Life Technologies Corporation. The transfer of this product is contingent on the buyer using the purchase product solely in research, excluding contract research or any fee for service research, and the buyer must not sell or otherwise transfer this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; (c) manufacturing or quality assurance or quality control, or (d) resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad CA 92008 USA or outlicensing@thermofisher.com
    • 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
      Flow CytometryNeat
    • Application NameYesNoMin DilutionMax Dilution
      Flow Cytometry1/51/200
      Functional Assays
      Immunofluorescence
      Western Blotting
    • Application NameYesNoMin DilutionMax Dilution
      Flow CytometryNeat1/10
    • Application NameYesNoMin DilutionMax Dilution
      Flow Cytometry1/501/200
      Immunofluorescence
      Western Blotting

    • Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. It is recommended that the user titrates the antibody for use in their own system to a concentration equivalent to their test reagent.
    • 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 106 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 CD335 Antibody Formats

    Formats Clone Applications Sizes available
    CD335 Antibody : Alexa Fluor® 488 AKS1 F 100 Tests/1ml
    CD335 Antibody : Purified AKS1 F, IF, WB 0.1 mg
    CD335 Antibody : Low Endotoxin AKS1 F, FN, IF, WB 0.5 mg
    CD335 Antibody : RPE AKS1 F 100 Tests
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

    Recommended Secondary Antibody

      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed)STAR117A0.5 mgE, WB
      STAR117A
      Goat anti Mouse IgG/A/M:Alk. Phos.STAR87A1 mgC, E, WB
      STAR87A
      Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed)STAR117D488GA0.1 mgF, IF
      STAR117D488GA
      Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed)STAR117D549GA0.1 mgF, IF
      STAR117D549GA
      Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)STAR117D649GA0.1 mgF, IF
      STAR117D649GA
      Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed)STAR117D680GA0.1 mgF, WB
      STAR117D680GA
      Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed)STAR117D800GA0.1 mgF, IF, WB
      STAR117D800GA
      Rabbit F(ab')2 anti Mouse IgG:Dylight®800STAR8D800GA0.1 mgF, IF, WB
      STAR8D800GA
      Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)STAR117F0.5 mgF
      STAR117F
      Goat anti Mouse IgG:FITC (Rat Adsorbed)STAR700.5 mgF
      STAR70
      Goat anti Mouse IgG (Fc):FITCSTAR120F1 mgC, F
      STAR120F
      Rabbit F(ab')2 anti Mouse IgG:FITCSTAR9B1 mgF
      STAR9B
      Human anti Mouse IgG1:HRPHCA036P0.1 mgE
      HCA036P
      Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)STAR117P0.5 mgE, WB
      STAR117P
      Goat anti Mouse IgG:HRP (Rat Adsorbed)STAR770.5 mgC, E, P
      STAR77
      Goat anti Mouse IgG (Fc):HRPSTAR120P1 mgE, WB
      STAR120P
      Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
      STAR13B
      Goat anti Mouse IgG/A/M:HRP (Human Adsorbed)STAR87P1 mgE
      STAR87P
      Rabbit F(ab')2 anti Mouse IgG:RPESTAR12A1 mlF
      STAR12A
      Goat anti Mouse IgG:RPE (Rat Adsorbed)STAR761 mlF
      STAR76
      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
      Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed)STAR117A0.5 mgE, WB
      STAR117A
      Goat anti Mouse IgG/A/M:Alk. Phos.STAR87A1 mgC, E, WB
      STAR87A
      Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed)STAR117D488GA0.1 mgF, IF
      STAR117D488GA
      Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed)STAR117D549GA0.1 mgF, IF
      STAR117D549GA
      Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed)STAR117D649GA0.1 mgF, IF
      STAR117D649GA
      Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed)STAR117D680GA0.1 mgF, WB
      STAR117D680GA
      Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed)STAR117D800GA0.1 mgF, IF, WB
      STAR117D800GA
      Rabbit F(ab')2 anti Mouse IgG:Dylight®800STAR8D800GA0.1 mgF, IF, WB
      STAR8D800GA
      Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed)STAR117F0.5 mgF
      STAR117F
      Goat anti Mouse IgG:FITC (Rat Adsorbed)STAR700.5 mgF
      STAR70
      Goat anti Mouse IgG (Fc):FITCSTAR120F1 mgC, F
      STAR120F
      Rabbit F(ab')2 anti Mouse IgG:FITCSTAR9B1 mgF
      STAR9B
      Human anti Mouse IgG1:HRPHCA036P0.1 mgE
      HCA036P
      Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed)STAR117P0.5 mgE, WB
      STAR117P
      Goat anti Mouse IgG:HRP (Rat Adsorbed)STAR770.5 mgC, E, P
      STAR77
      Goat anti Mouse IgG (Fc):HRPSTAR120P1 mgE, WB
      STAR120P
      Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed)STAR13B1 mgC, E, P, RE, WB
      STAR13B
      Goat anti Mouse IgG/A/M:HRP (Human Adsorbed)STAR87P1 mgE
      STAR87P
      Rabbit F(ab')2 anti Mouse IgG:RPESTAR12A1 mlF
      STAR12A
      Goat anti Mouse IgG:RPE (Rat Adsorbed)STAR761 mlF
      STAR76

      Recommended Negative Isotype Control

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative Control:RPEMCA928PE100 TestsF
        MCA928PE
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative Control:Low EndotoxinMCA928EL0.5 mgC, F
        MCA928EL
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative Control:Alexa Fluor® 488MCA928A488100 Tests/1mlF
        MCA928A488
        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse IgG1 Negative ControlMCA928100 TestsF
        MCA928

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              References

              Further Reading

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