alkaline phosphatase labeled goat anti mouse igg1  (SouthernBiotech)

 
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    Name:
    Goat F ab 2 Anti Mouse Kappa BIOT
    Description:

    Catalog Number:
    1052-08
    Price:
    None
    Source:
    Pepsin digest of Goat Anti-Mouse Kappa (SB Cat. No. 1050)
    Applications:
    Quality tested applications for relevant formats include -ELISA 1Flow CytometryOther referenced applications for relevant formats include -Immunohistochemistry-Paraffin Sections 8ELISpot 1 Stimulation 2-7
    Format:
    BIOT (Biotin)
    Isotype:
    Goat F(ab')2 IgG
    Buy from Supplier


    Structured Review

    SouthernBiotech alkaline phosphatase labeled goat anti mouse igg1
    Goat F ab 2 Anti Mouse Kappa BIOT

    https://www.bioz.com/result/alkaline phosphatase labeled goat anti mouse igg1/product/SouthernBiotech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase labeled goat anti mouse igg1 - by Bioz Stars, 2021-05
    94/100 stars

    Images

    1) Product Images from "VH3 Antibody Response to Immunization with Pneumococcal Polysaccharide Vaccine in Middle-Aged and Elderly Persons ▿"

    Article Title: VH3 Antibody Response to Immunization with Pneumococcal Polysaccharide Vaccine in Middle-Aged and Elderly Persons ▿

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00408-10

    IgG antibody response.
    Figure Legend Snippet: IgG antibody response.

    Techniques Used:

    2) Product Images from "Activating an adaptive immune response from a hydrogel scaffold imparts regenerative wound healing."

    Article Title: Activating an adaptive immune response from a hydrogel scaffold imparts regenerative wound healing.

    Journal: Nature materials

    doi: 10.1038/s41563-020-00844-w

    D-MAP induces antibody responses and recruitment of myeloid cells via adaptive immunity. a-c ) Measurement of anti-D specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. d-f ) Measurement of anti-L specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. Each data point represents one animal and all analysis in a-f is by unpaired two-tailed t-test comparing each condition to L only. g-i ) Measurement of anti-D specific IgG subtype antibodies in Balb/c or Balb/c.Rag2 −/− γc −/− mice given a subcutaneous injection of D-MAP 21 days after injection. Each data point represents one animal and all analysis in g-I is by unpaired two-tailed t-test (** denotes p=0.0022). j-l ) Representative examples of confocal immunofluorescent imaging for CD11b, DAPI, and hydrogel from subcutaneous implants of L- or D-MAP hydrogel implants in Balb/c or Balb/c.Rag2 −/− γc −/− mice. Scale = 200μm. (j) and quantification of total DAPI + cells (k) and CD11b + myeloid cells (l). Data is plotted as a scatter plot showing the mean and standard deviation. Each point represents average of 3 slides for each wound. All analysis is by unpaired two-tailed t-test (* denotes p=0.0455, *** denotes p=0.0006, **** denotes p
    Figure Legend Snippet: D-MAP induces antibody responses and recruitment of myeloid cells via adaptive immunity. a-c ) Measurement of anti-D specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. d-f ) Measurement of anti-L specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. Each data point represents one animal and all analysis in a-f is by unpaired two-tailed t-test comparing each condition to L only. g-i ) Measurement of anti-D specific IgG subtype antibodies in Balb/c or Balb/c.Rag2 −/− γc −/− mice given a subcutaneous injection of D-MAP 21 days after injection. Each data point represents one animal and all analysis in g-I is by unpaired two-tailed t-test (** denotes p=0.0022). j-l ) Representative examples of confocal immunofluorescent imaging for CD11b, DAPI, and hydrogel from subcutaneous implants of L- or D-MAP hydrogel implants in Balb/c or Balb/c.Rag2 −/− γc −/− mice. Scale = 200μm. (j) and quantification of total DAPI + cells (k) and CD11b + myeloid cells (l). Data is plotted as a scatter plot showing the mean and standard deviation. Each point represents average of 3 slides for each wound. All analysis is by unpaired two-tailed t-test (* denotes p=0.0455, *** denotes p=0.0006, **** denotes p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay, Two Tailed Test, Injection, Imaging, Standard Deviation

    3) Product Images from "A Monoclonal Antibody to Cryptococcus neoformans Glucuronoxylomannan Manifests Hydrolytic Activity for Both Peptides and Polysaccharides *"

    Article Title: A Monoclonal Antibody to Cryptococcus neoformans Glucuronoxylomannan Manifests Hydrolytic Activity for Both Peptides and Polysaccharides *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.767582

    mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C IgG1 control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.
    Figure Legend Snippet: mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C IgG1 control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.

    Techniques Used: Flow Cytometry, Cytometry, Construct, Incubation

    4) Product Images from "Activating an adaptive immune response from a hydrogel scaffold imparts regenerative wound healing"

    Article Title: Activating an adaptive immune response from a hydrogel scaffold imparts regenerative wound healing

    Journal: bioRxiv

    doi: 10.1101/2020.05.27.117317

    D-MAP induces antibody responses in vivo and requires an intact adaptive immunity for optimal myeloid cell recruitment. a-c) Measurement of anti-D specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. d-f) Measurement of anti-L specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. g-i) Measurement of anti-D specific IgG subtype antibodies in Balb/c or Balb/c.Rag2 −/− γc −/− mice given a subcutaneous injection of D-MAP 21 days after injection. j-l) Representative examples of confocal immunofluorescent imaging for CD11b, DAPI, and hydrogel from subcutaneous implants of L- or D-MAP hydrogel implants in Balb/c or Balb/c.Rag2 −/− γc −/− mice (j) and quantification of total DAPI+ cells (k), CD11b + myeloid cells (l). Data is plotted as a scatter plot showing the mean and standard deviation. *, **, *** represent statistical significance by student t-test for the comparison indicated.
    Figure Legend Snippet: D-MAP induces antibody responses in vivo and requires an intact adaptive immunity for optimal myeloid cell recruitment. a-c) Measurement of anti-D specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. d-f) Measurement of anti-L specific IgG subtype antibodies by ELISA 21 days following wound healing experiments in SKH1 mice treated with indicated hydrogels. g-i) Measurement of anti-D specific IgG subtype antibodies in Balb/c or Balb/c.Rag2 −/− γc −/− mice given a subcutaneous injection of D-MAP 21 days after injection. j-l) Representative examples of confocal immunofluorescent imaging for CD11b, DAPI, and hydrogel from subcutaneous implants of L- or D-MAP hydrogel implants in Balb/c or Balb/c.Rag2 −/− γc −/− mice (j) and quantification of total DAPI+ cells (k), CD11b + myeloid cells (l). Data is plotted as a scatter plot showing the mean and standard deviation. *, **, *** represent statistical significance by student t-test for the comparison indicated.

    Techniques Used: In Vivo, Enzyme-linked Immunosorbent Assay, Mouse Assay, Injection, Imaging, Standard Deviation

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    Sandwich ELISA:

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    Article Snippet: .. Titers of IgG in serum were determined by sandwich ELISA using specific goat anti–mouse polyclonal reagents (Southern Biotechnology Associates, Inc.). .. Purified IgG was used for the standard curve and for calculation of Ab concentration.

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    Enzyme-linked Immunosorbent Assay:

    Article Title: Selection at Multiple Checkpoints Focuses VH12 B Cell Differentiation toward a Single B-1 Cell Specificity
    Article Snippet: Therefore, we cotransfected into P3-X63-Ag8.653 myeloma cells with the 10/G4 or 2-12 expression constructs with L chain expression vectors containing Vκ4/5H or Vκ21C rearrangements as described . .. To test whether a complete Ig molecule was formed, supernatant was subjected to ELISA using microtiter plates coated with polyclonal goat anti–mouse μ (Southern Biotechnology Associates, Inc.) and alkaline phosphatase–labeled polyclonal goat anti–mouse κ (Southern Biotechnology Associates, Inc.) to develop the reaction. .. In those cases where Ig secretion was not detected, the production of H and L chains was confirmed by ELISA using cell lysates and the polyclonal goat anti–mouse μ– or polyclonal goat anti–mouse κ-coated plates as above.

    Article Title: Selection at Multiple Checkpoints Focuses VH12 B Cell Differentiation toward a Single B-1 Cell Specificity
    Article Snippet: To test whether a complete Ig molecule was formed, supernatant was subjected to ELISA using microtiter plates coated with polyclonal goat anti–mouse μ (Southern Biotechnology Associates, Inc.) and alkaline phosphatase–labeled polyclonal goat anti–mouse κ (Southern Biotechnology Associates, Inc.) to develop the reaction. .. In those cases where Ig secretion was not detected, the production of H and L chains was confirmed by ELISA using cell lysates and the polyclonal goat anti–mouse μ– or polyclonal goat anti–mouse κ-coated plates as above. ..

    Expressing:

    Article Title: CXCR5 CAR-T cells simultaneously target B cell non-Hodgkin’s lymphoma and tumor-supportive follicular T helper cells
    Article Snippet: 2) Monocytes and DCs: FITC anti-CD1c (L161, #331518, 1:100), PE anti-CXCR5 (51505, #FAB190P-100, R & D Systems/Bio-Techne, 1:10), PE/Cy7 anti-CD11c (Bu15, #337216, 1:100), APC anti-CD14 (HCD14, #325608, 1:100), APC/Fire750 anti-CD303 (201A, #354236, 1:100), BV421 anti-HLA-DR (L243, #307636, 1:100), BV510 anti-CD19 (SJ25C1, #363020, 1:100) and BV510 anti-CD3 (SK7, #344828, 1:100). .. CAR expression on transduced mouse splenocytes was detected using polyclonal PE-labeled goat anti-mouse IgG-Ab (#1030-09, Southern Biotech/Biozol, 1:200). ..

    Staining:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

    Modification:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

    Avidin-Biotin Assay:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

    Blocking Assay:

    Article Title: A murine Ig light chain transgene reveals IGKV3 gene contributions to anti-collagen types IV and II specificities
    Article Snippet: .. Sections of kidneys frozen in OCT medium were stained for mouse kappa Ig and images acquired essentially as described (direct IF) , with the following modification: sections were blocked with the avidin-biotin blocking kit (Life Technologies, Frederick, MD, USA), and deposited mLCV3-Tg detected using biotin-conjugated goat-anti-mouse-Ig-kappa (Southern Biotech) and streptavidin-Alexa Fluor 488 (Life Technologies). .. For indirect IF, sections of kidney from NOD-scid-gamma immunodeficient mice (Jackson Laboratory) were incubated with supernatant from mLCV3-Tg/kKO mouse splenocytes cultured with R848 + CpG as described above, or with positive control mouse anti-alpha3(IV)NC1 IgG MAB3 (Eurodiagnostica), and mouse kappa detected as described above.

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  • 94
    SouthernBiotech alkaline phosphatase labeled goat anti mouse igg1
    mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C <t>IgG1</t> control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.
    Alkaline Phosphatase Labeled Goat Anti Mouse Igg1, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alkaline phosphatase labeled goat anti mouse igg1/product/SouthernBiotech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase labeled goat anti mouse igg1 - by Bioz Stars, 2021-05
    94/100 stars
      Buy from Supplier

    93
    SouthernBiotech ap labelled goat anti mouse igg1
    Expression of haemoglobin in Nippostrongylus brasiliensis . A ) N. brasilensis adult worms isolated from mouse guts were formalin fixed and stained with 4E8g (Anti-Hb). Detection was carried out using the anti-mouse <t>IgG</t> detection system with DAB. Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. 400× magnification. B ) Small intestines of mice infected with N. brasiliensis (Day 7 post-infection) were formalin fixed, stained with Anti-Hb and counterstained with haemotoxylin. N. brasiliensis worms are indicated by arrows. 40× magnification. C ) N.brasilensis larvae in mouse intestines stained with 4E8g (left) and isotype control antibody (mouse <t>IgG1)(right).</t>
    Ap Labelled Goat Anti Mouse Igg1, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ap labelled goat anti mouse igg1/product/SouthernBiotech
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ap labelled goat anti mouse igg1 - by Bioz Stars, 2021-05
    93/100 stars
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    94
    SouthernBiotech goat anti mouse igg2a
    Generation of dsRNA in tumor cells transfected with pSIN-β . (A) . RT-PCR confirmed the presence of sindbis virus nsp4 gene mRNA and its anti-sense strand in tumor cells transfected with pSIN-β. TC-1 cells were transfected with pCMV-β (pCMV) or pSin-β (pSIN), or left untreated (N/A). Total RNA was reverse transcribed into DNA with oligo dT primer or primers specific to the nsp4 gene (forward p4F or reverse p4R) before PCR amplification. This experiment was repeated twice with similar results. (B) . ELISA confirmed the presence of an elevated level of dsRNA in TC-1 cells transfected with pSIN-β (n = 3). Total dsRNA was isolated from TC-1 cells transfected with pCMV-β or pSIN-β and used to coat ELISA plate. The primary Ab was the J2 anti-dsRNA <t>IgG2a.</t> *, p = 0.004. (C) . Transfection of pSIN-β into TC-1 cells inhibited cell growth. TC-1 cells (20 000 cells/well) were transfected with the same amount (0.4 μg) of pCMV-β, pSIN-β, or pSIN-β-Δnsp (n = 4). Cell numbers were quantified using MTT assay and normalized to cells treated with sterile PBS. Data shown are mean ± S.E.M. **, at 48 and 72 h, the value of the pSIN-β were different from that of the pCMV-β and the pSIN-β-Δnsp (p
    Goat Anti Mouse Igg2a, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/goat anti mouse igg2a/product/SouthernBiotech
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    goat anti mouse igg2a - by Bioz Stars, 2021-05
    94/100 stars
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    86
    SouthernBiotech alkaline phosphatase conjugated goat anti mouse igg
    Anti-HEL autoantibodies enhance accumulation of HEL-specific CD4+ T cells. Splenocytes from TCR+ mice (bearing the CD45.1 allelic marker) were labeled with the cell division dye CFSE and injected intravenously into 24 HEL+ recipients (CD45.2) on day 0. Twelve recipients were given 200 μL HEL-immune serum and twelve were given 200 μL control serum i.p. starting on day −1 and on alternate days thereafter in surviving mice. Flow cytometry on recipient spleens was performed on days 3, 5, 7, and 10, each time using three mice from each treatment group plus two control nontransgenic mice that had received cells but no serum injections. A : Representative flow cytometric plots (from day 5 after transfer) showing gates used to track donor-derived CD4 + cells and B220 + “tracer” B cells ( upper panel ), plus TCR HEL expression and CFSE dilution on the donor-derived CD4 + cells ( lower panel ). B : Graphs show the ratio of donor CD4 + cells to donor B220 + cells, and the shaded areas represent the proportion of TCR HEL + cells among donor-derived CD4 + cells. C : Percentage of CFSE-diluted cells among donor-derived CD4 + cells. In ( B ) and ( C ), symbols indicate group means; error bars show the SD. Statistical differences between HEL-immune and control serum groups were analyzed using a ratio t test (see research design and methods ). The data shown are from one of three experiments that yielded comparable results. D : The experiment described in A – C was repeated, except that the TCR+ donors were CD45.2+ Bim-deficient mice and the recipients were CD45.1+. Histogram ( top ) displays CFSE on donor (CD45.2+) CD4+ cells in HEL+ hosts that received HEL-immune or control serum, or in nontransgenic hosts as indicated, with a summary of the data ( bottom ) for multiple recipients. E : Diabetes incidence in TCR+HEL+ neonates injected on days 2, 4, and 6 after birth with serum from HEL-immunized mice and on days 1, 3, 5, 8, 11, 14, and 17 after birth with Fcγ R -blocking antibody or rat <t>IgG2b</t> isotype control. Statistical analysis used the log-rank method.
    Alkaline Phosphatase Conjugated Goat Anti Mouse Igg, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alkaline phosphatase conjugated goat anti mouse igg/product/SouthernBiotech
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alkaline phosphatase conjugated goat anti mouse igg - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    Image Search Results


    mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C IgG1 control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.

    Journal: The Journal of Biological Chemistry

    Article Title: A Monoclonal Antibody to Cryptococcus neoformans Glucuronoxylomannan Manifests Hydrolytic Activity for Both Peptides and Polysaccharides *

    doi: 10.1074/jbc.M116.767582

    Figure Lengend Snippet: mAb 18B7 modifies the capsule resulting in more 18B7 reactivity. A , flow cytometry gating strategy for Uvitex2b-positive cells, > 99% ( left plot ), used to construct histograms of 18B7-Alexa-568 reactivity cryptococcal capsules following 39 days of incubation with 0 (control), 1, 10, or 50 μg/ml mAb 18B7 (unlabeled) and 50 μg/ml MOPC-31C IgG1 control. Histograms shown are from one representative experiment. B , micrograph representations of samples analyzed in A , showing cell wall and capsule stainings. India ink counterstain was used to visualized the capsule under bright field. Scale bar, 10 μm.

    Article Snippet: Completion of the assay was performed by successively adding the anti-GXM IgG1 mAb 18B7 at 10 μg/ml and then alkaline phosphatase-labeled goat anti-mouse IgG1 at 1 μg/ml (SouthernBiotech).

    Techniques: Flow Cytometry, Cytometry, Construct, Incubation

    Expression of haemoglobin in Nippostrongylus brasiliensis . A ) N. brasilensis adult worms isolated from mouse guts were formalin fixed and stained with 4E8g (Anti-Hb). Detection was carried out using the anti-mouse IgG detection system with DAB. Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. 400× magnification. B ) Small intestines of mice infected with N. brasiliensis (Day 7 post-infection) were formalin fixed, stained with Anti-Hb and counterstained with haemotoxylin. N. brasiliensis worms are indicated by arrows. 40× magnification. C ) N.brasilensis larvae in mouse intestines stained with 4E8g (left) and isotype control antibody (mouse IgG1)(right).

    Journal: PLoS Neglected Tropical Diseases

    Article Title: A Cross-Reactive Monoclonal Antibody to Nematode Haemoglobin Enhances Protective Immune Responses to Nippostrongylus brasiliensis

    doi: 10.1371/journal.pntd.0002395

    Figure Lengend Snippet: Expression of haemoglobin in Nippostrongylus brasiliensis . A ) N. brasilensis adult worms isolated from mouse guts were formalin fixed and stained with 4E8g (Anti-Hb). Detection was carried out using the anti-mouse IgG detection system with DAB. Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. 400× magnification. B ) Small intestines of mice infected with N. brasiliensis (Day 7 post-infection) were formalin fixed, stained with Anti-Hb and counterstained with haemotoxylin. N. brasiliensis worms are indicated by arrows. 40× magnification. C ) N.brasilensis larvae in mouse intestines stained with 4E8g (left) and isotype control antibody (mouse IgG1)(right).

    Article Snippet: For the detection of haemoglobin-specific mouse IgG and IgE, serum collected from Anisakis -infected mice in a previous study was used with AP-labelled goat anti-mouse IgG1 (Southern Biotech, USA) and goat anti-mouse IgE (Southern Biotech, USA) as secondary antibodies.

    Techniques: Expressing, Isolation, Staining, Mouse Assay, Infection

    Expression of haemoglobin in Ascaris lumbricoides . A ) Longitudinal sections and B ) cross sections of a formalin-fixed A. lumbricodes adult worm was stained with 4E8g (Anti-Hb) and detected with the anti-mouse IgG detection system with DAB. C ) A.lumbricoides section stained with isotype control antibody (mouse IgG1). Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. All photos taken at 40× magnification.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: A Cross-Reactive Monoclonal Antibody to Nematode Haemoglobin Enhances Protective Immune Responses to Nippostrongylus brasiliensis

    doi: 10.1371/journal.pntd.0002395

    Figure Lengend Snippet: Expression of haemoglobin in Ascaris lumbricoides . A ) Longitudinal sections and B ) cross sections of a formalin-fixed A. lumbricodes adult worm was stained with 4E8g (Anti-Hb) and detected with the anti-mouse IgG detection system with DAB. C ) A.lumbricoides section stained with isotype control antibody (mouse IgG1). Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. All photos taken at 40× magnification.

    Article Snippet: For the detection of haemoglobin-specific mouse IgG and IgE, serum collected from Anisakis -infected mice in a previous study was used with AP-labelled goat anti-mouse IgG1 (Southern Biotech, USA) and goat anti-mouse IgE (Southern Biotech, USA) as secondary antibodies.

    Techniques: Expressing, Staining

    Immunoblotting with purified haemoglobin. A ) Specific IgG1 against purified Anisakis haemoglobin in serum from mice infected at week 0 with two live A. pegreffii L3 and re-infected at week 8 with 2 L3. Serum was collected at week 11. Each lane represents one individual mouse. B ) Specific IgE against purified Anisakis haemoglobin in mouse serum, collected as in (A). Each lane represents one individual mouse. C ) Specific IgG against purified Ascaris haemoglobin in human serum samples that were negative (0 kU/L specific IgE, CAP-RAST) or positive (0.9–17.9 kU/L specific IgE, CAP-RAST) for Ascaris specific antibodies. Each lane represents one individual.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: A Cross-Reactive Monoclonal Antibody to Nematode Haemoglobin Enhances Protective Immune Responses to Nippostrongylus brasiliensis

    doi: 10.1371/journal.pntd.0002395

    Figure Lengend Snippet: Immunoblotting with purified haemoglobin. A ) Specific IgG1 against purified Anisakis haemoglobin in serum from mice infected at week 0 with two live A. pegreffii L3 and re-infected at week 8 with 2 L3. Serum was collected at week 11. Each lane represents one individual mouse. B ) Specific IgE against purified Anisakis haemoglobin in mouse serum, collected as in (A). Each lane represents one individual mouse. C ) Specific IgG against purified Ascaris haemoglobin in human serum samples that were negative (0 kU/L specific IgE, CAP-RAST) or positive (0.9–17.9 kU/L specific IgE, CAP-RAST) for Ascaris specific antibodies. Each lane represents one individual.

    Article Snippet: For the detection of haemoglobin-specific mouse IgG and IgE, serum collected from Anisakis -infected mice in a previous study was used with AP-labelled goat anti-mouse IgG1 (Southern Biotech, USA) and goat anti-mouse IgE (Southern Biotech, USA) as secondary antibodies.

    Techniques: Purification, Mouse Assay, Infection, RAST Test

    Expression of haemoglobin in Anisakis pegreffii . A–D ) A. pegreffii L3 collected from Thyrsites atun were formalin fixed and stained with 4E8g (Anti-Hb). E–F ) A. pegreffii L3 sections were stained with isotype control antibody (mouse IgG1). Detection was carried out using the anti-mouse IgG detection system with DAB. Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. All photos taken at 200× magnification.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: A Cross-Reactive Monoclonal Antibody to Nematode Haemoglobin Enhances Protective Immune Responses to Nippostrongylus brasiliensis

    doi: 10.1371/journal.pntd.0002395

    Figure Lengend Snippet: Expression of haemoglobin in Anisakis pegreffii . A–D ) A. pegreffii L3 collected from Thyrsites atun were formalin fixed and stained with 4E8g (Anti-Hb). E–F ) A. pegreffii L3 sections were stained with isotype control antibody (mouse IgG1). Detection was carried out using the anti-mouse IgG detection system with DAB. Haemoglobin is stained in brown. Counterstaining was performed with haemotoxylin. All photos taken at 200× magnification.

    Article Snippet: For the detection of haemoglobin-specific mouse IgG and IgE, serum collected from Anisakis -infected mice in a previous study was used with AP-labelled goat anti-mouse IgG1 (Southern Biotech, USA) and goat anti-mouse IgE (Southern Biotech, USA) as secondary antibodies.

    Techniques: Expressing, Staining

    Generation of dsRNA in tumor cells transfected with pSIN-β . (A) . RT-PCR confirmed the presence of sindbis virus nsp4 gene mRNA and its anti-sense strand in tumor cells transfected with pSIN-β. TC-1 cells were transfected with pCMV-β (pCMV) or pSin-β (pSIN), or left untreated (N/A). Total RNA was reverse transcribed into DNA with oligo dT primer or primers specific to the nsp4 gene (forward p4F or reverse p4R) before PCR amplification. This experiment was repeated twice with similar results. (B) . ELISA confirmed the presence of an elevated level of dsRNA in TC-1 cells transfected with pSIN-β (n = 3). Total dsRNA was isolated from TC-1 cells transfected with pCMV-β or pSIN-β and used to coat ELISA plate. The primary Ab was the J2 anti-dsRNA IgG2a. *, p = 0.004. (C) . Transfection of pSIN-β into TC-1 cells inhibited cell growth. TC-1 cells (20 000 cells/well) were transfected with the same amount (0.4 μg) of pCMV-β, pSIN-β, or pSIN-β-Δnsp (n = 4). Cell numbers were quantified using MTT assay and normalized to cells treated with sterile PBS. Data shown are mean ± S.E.M. **, at 48 and 72 h, the value of the pSIN-β were different from that of the pCMV-β and the pSIN-β-Δnsp (p

    Journal: BMC Cancer

    Article Title: Replicase-based plasmid DNA shows anti-tumor activity

    doi: 10.1186/1471-2407-11-110

    Figure Lengend Snippet: Generation of dsRNA in tumor cells transfected with pSIN-β . (A) . RT-PCR confirmed the presence of sindbis virus nsp4 gene mRNA and its anti-sense strand in tumor cells transfected with pSIN-β. TC-1 cells were transfected with pCMV-β (pCMV) or pSin-β (pSIN), or left untreated (N/A). Total RNA was reverse transcribed into DNA with oligo dT primer or primers specific to the nsp4 gene (forward p4F or reverse p4R) before PCR amplification. This experiment was repeated twice with similar results. (B) . ELISA confirmed the presence of an elevated level of dsRNA in TC-1 cells transfected with pSIN-β (n = 3). Total dsRNA was isolated from TC-1 cells transfected with pCMV-β or pSIN-β and used to coat ELISA plate. The primary Ab was the J2 anti-dsRNA IgG2a. *, p = 0.004. (C) . Transfection of pSIN-β into TC-1 cells inhibited cell growth. TC-1 cells (20 000 cells/well) were transfected with the same amount (0.4 μg) of pCMV-β, pSIN-β, or pSIN-β-Δnsp (n = 4). Cell numbers were quantified using MTT assay and normalized to cells treated with sterile PBS. Data shown are mean ± S.E.M. **, at 48 and 72 h, the value of the pSIN-β were different from that of the pCMV-β and the pSIN-β-Δnsp (p

    Article Snippet: Horseradish peroxidase (HRP) labeled goat anti-mouse IgG2a (5 000-fold dilution Southern Biotechnology Associates, Birmingham, AL) was added to the wells, followed by 1 h of incubation at 37°C.

    Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification, Enzyme-linked Immunosorbent Assay, Isolation, MTT Assay

    Anti-HEL autoantibodies enhance accumulation of HEL-specific CD4+ T cells. Splenocytes from TCR+ mice (bearing the CD45.1 allelic marker) were labeled with the cell division dye CFSE and injected intravenously into 24 HEL+ recipients (CD45.2) on day 0. Twelve recipients were given 200 μL HEL-immune serum and twelve were given 200 μL control serum i.p. starting on day −1 and on alternate days thereafter in surviving mice. Flow cytometry on recipient spleens was performed on days 3, 5, 7, and 10, each time using three mice from each treatment group plus two control nontransgenic mice that had received cells but no serum injections. A : Representative flow cytometric plots (from day 5 after transfer) showing gates used to track donor-derived CD4 + cells and B220 + “tracer” B cells ( upper panel ), plus TCR HEL expression and CFSE dilution on the donor-derived CD4 + cells ( lower panel ). B : Graphs show the ratio of donor CD4 + cells to donor B220 + cells, and the shaded areas represent the proportion of TCR HEL + cells among donor-derived CD4 + cells. C : Percentage of CFSE-diluted cells among donor-derived CD4 + cells. In ( B ) and ( C ), symbols indicate group means; error bars show the SD. Statistical differences between HEL-immune and control serum groups were analyzed using a ratio t test (see research design and methods ). The data shown are from one of three experiments that yielded comparable results. D : The experiment described in A – C was repeated, except that the TCR+ donors were CD45.2+ Bim-deficient mice and the recipients were CD45.1+. Histogram ( top ) displays CFSE on donor (CD45.2+) CD4+ cells in HEL+ hosts that received HEL-immune or control serum, or in nontransgenic hosts as indicated, with a summary of the data ( bottom ) for multiple recipients. E : Diabetes incidence in TCR+HEL+ neonates injected on days 2, 4, and 6 after birth with serum from HEL-immunized mice and on days 1, 3, 5, 8, 11, 14, and 17 after birth with Fcγ R -blocking antibody or rat IgG2b isotype control. Statistical analysis used the log-rank method.

    Journal: Diabetes

    Article Title: Anti-Islet Autoantibodies Trigger Autoimmune Diabetes in the Presence of an Increased Frequency of Islet-Reactive CD4 T Cells

    doi: 10.2337/db10-1344

    Figure Lengend Snippet: Anti-HEL autoantibodies enhance accumulation of HEL-specific CD4+ T cells. Splenocytes from TCR+ mice (bearing the CD45.1 allelic marker) were labeled with the cell division dye CFSE and injected intravenously into 24 HEL+ recipients (CD45.2) on day 0. Twelve recipients were given 200 μL HEL-immune serum and twelve were given 200 μL control serum i.p. starting on day −1 and on alternate days thereafter in surviving mice. Flow cytometry on recipient spleens was performed on days 3, 5, 7, and 10, each time using three mice from each treatment group plus two control nontransgenic mice that had received cells but no serum injections. A : Representative flow cytometric plots (from day 5 after transfer) showing gates used to track donor-derived CD4 + cells and B220 + “tracer” B cells ( upper panel ), plus TCR HEL expression and CFSE dilution on the donor-derived CD4 + cells ( lower panel ). B : Graphs show the ratio of donor CD4 + cells to donor B220 + cells, and the shaded areas represent the proportion of TCR HEL + cells among donor-derived CD4 + cells. C : Percentage of CFSE-diluted cells among donor-derived CD4 + cells. In ( B ) and ( C ), symbols indicate group means; error bars show the SD. Statistical differences between HEL-immune and control serum groups were analyzed using a ratio t test (see research design and methods ). The data shown are from one of three experiments that yielded comparable results. D : The experiment described in A – C was repeated, except that the TCR+ donors were CD45.2+ Bim-deficient mice and the recipients were CD45.1+. Histogram ( top ) displays CFSE on donor (CD45.2+) CD4+ cells in HEL+ hosts that received HEL-immune or control serum, or in nontransgenic hosts as indicated, with a summary of the data ( bottom ) for multiple recipients. E : Diabetes incidence in TCR+HEL+ neonates injected on days 2, 4, and 6 after birth with serum from HEL-immunized mice and on days 1, 3, 5, 8, 11, 14, and 17 after birth with Fcγ R -blocking antibody or rat IgG2b isotype control. Statistical analysis used the log-rank method.

    Article Snippet: Serial serum dilutions were applied followed by alkaline phosphatase-conjugated goat anti-mouse IgG or IgG2a (Southern Biotechnology) and then Phosphatase Substrate (Sigma).

    Techniques: Mouse Assay, Marker, Labeling, Injection, Flow Cytometry, Cytometry, Derivative Assay, Expressing, Blocking Assay

    Effect of passive transfer of antibody on diabetes in TCR+ HEL+ mice. A : Breeding strategy for maternal antibody transfer experiments. Nontransgenic B10.BR females were immunized with HEL or OVA protein emulsified in CFA to induce specific antibodies and then mated with TCR+HEL+ males to yield 25% TCR+HEL+ offspring and equal numbers of singly transgenic or nontransgenic littermates. B : Anti-HEL IgG titers in newborns from mothers immunized as illustrated in ( A ); 70-day-old pups from immunized or unimmunized mothers and mothers 5 weeks after immunization. C : Diabetes incidence in offspring with the indicated genotypes bred from HEL-immunized mothers. D : Diabetes incidence in TCR+HEL+ offspring of mothers immunized with either HEL (closed symbols) or OVA (open symbols). E and F : Diabetes incidence in TCR+HEL+ neonates injected on days 1, 3, and 5 after birth with serum ( E ) or purified IgG ( F ) from HEL- or OVA-immunized mice.

    Journal: Diabetes

    Article Title: Anti-Islet Autoantibodies Trigger Autoimmune Diabetes in the Presence of an Increased Frequency of Islet-Reactive CD4 T Cells

    doi: 10.2337/db10-1344

    Figure Lengend Snippet: Effect of passive transfer of antibody on diabetes in TCR+ HEL+ mice. A : Breeding strategy for maternal antibody transfer experiments. Nontransgenic B10.BR females were immunized with HEL or OVA protein emulsified in CFA to induce specific antibodies and then mated with TCR+HEL+ males to yield 25% TCR+HEL+ offspring and equal numbers of singly transgenic or nontransgenic littermates. B : Anti-HEL IgG titers in newborns from mothers immunized as illustrated in ( A ); 70-day-old pups from immunized or unimmunized mothers and mothers 5 weeks after immunization. C : Diabetes incidence in offspring with the indicated genotypes bred from HEL-immunized mothers. D : Diabetes incidence in TCR+HEL+ offspring of mothers immunized with either HEL (closed symbols) or OVA (open symbols). E and F : Diabetes incidence in TCR+HEL+ neonates injected on days 1, 3, and 5 after birth with serum ( E ) or purified IgG ( F ) from HEL- or OVA-immunized mice.

    Article Snippet: Serial serum dilutions were applied followed by alkaline phosphatase-conjugated goat anti-mouse IgG or IgG2a (Southern Biotechnology) and then Phosphatase Substrate (Sigma).

    Techniques: Mouse Assay, Transgenic Assay, Injection, Purification

    High autoantibodies and requirement for B cells for accelerated progression to diabetes in Roquin san/san TCR+HEL+ mice. A and B : Titers of anti-HEL IgG ( A ) and IgG2a ( B ) in TCR+HEL+ mice of the indicated Roquin genotypes and diabetes status. C : Anti-HEL IgG titers in diabetic TCR+HEL+ mice of the indicated genotypes. D : Diabetes incidence in Roquin san/san TCR+HEL+ mice that were either B cell deficient ( Cd79a ken/ken ; filled circles) or B cell sufficient ( Cd79a +/+ ; triangles) and in Roquin +/+ Cd79a ken/ken TCR+HEL+ mice (open circles). E : Diabetes incidence in Roquin san/san TCR+HEL+ mice with or without an IgMD HEL transgene that prevents isotype switching of HEL-specific B cells, and control TCR+HEL+ mice with normal Roquin and B-cell repertoire. F : Diabetes incidence in Roquin san/+ TCR+HEL+ mice after transfer of a single 200 μL dose of serum from diabetic Roquin san/san TCR+HEL+ mice (closed symbols) or from diabetic Roquin san/+ or Roquin +/+ TCR+HEL+ mice (open symbols).

    Journal: Diabetes

    Article Title: Anti-Islet Autoantibodies Trigger Autoimmune Diabetes in the Presence of an Increased Frequency of Islet-Reactive CD4 T Cells

    doi: 10.2337/db10-1344

    Figure Lengend Snippet: High autoantibodies and requirement for B cells for accelerated progression to diabetes in Roquin san/san TCR+HEL+ mice. A and B : Titers of anti-HEL IgG ( A ) and IgG2a ( B ) in TCR+HEL+ mice of the indicated Roquin genotypes and diabetes status. C : Anti-HEL IgG titers in diabetic TCR+HEL+ mice of the indicated genotypes. D : Diabetes incidence in Roquin san/san TCR+HEL+ mice that were either B cell deficient ( Cd79a ken/ken ; filled circles) or B cell sufficient ( Cd79a +/+ ; triangles) and in Roquin +/+ Cd79a ken/ken TCR+HEL+ mice (open circles). E : Diabetes incidence in Roquin san/san TCR+HEL+ mice with or without an IgMD HEL transgene that prevents isotype switching of HEL-specific B cells, and control TCR+HEL+ mice with normal Roquin and B-cell repertoire. F : Diabetes incidence in Roquin san/+ TCR+HEL+ mice after transfer of a single 200 μL dose of serum from diabetic Roquin san/san TCR+HEL+ mice (closed symbols) or from diabetic Roquin san/+ or Roquin +/+ TCR+HEL+ mice (open symbols).

    Article Snippet: Serial serum dilutions were applied followed by alkaline phosphatase-conjugated goat anti-mouse IgG or IgG2a (Southern Biotechnology) and then Phosphatase Substrate (Sigma).

    Techniques: Mouse Assay

    A maternally transmitted epigenetic factor increases diabetes incidence in the TCR+HEL+ offspring of formerly diabetic mothers. A : Diabetes incidence in TCR+HEL+ mice bred from TCR+HEL+ mothers or fathers that had spontaneously developed diabetes and had euglycemia stably restored by transplanting nontransgenic islets. The diabetes-cured TCR+HEL+ parents were bred with nontransgenic partners. Statistical analysis used the log-rank method. B : Diabetes incidence for the cohort described in ( A ) stratified by sex of the offspring. C : For each of 20 diabetes-cured TCR+HEL+ female breeders used to generate the cohort described in ( A ), two serial serum anti-HEL IgG titers (connected by a line) are plotted against the time of blood sampling relative to islet transplantation on day 0 ( left ). Serum anti-HEL IgG titers for 20 nondiabetic TCR+HEL+ females from the same colony as the diabetes-cured females ( right ).

    Journal: Diabetes

    Article Title: Anti-Islet Autoantibodies Trigger Autoimmune Diabetes in the Presence of an Increased Frequency of Islet-Reactive CD4 T Cells

    doi: 10.2337/db10-1344

    Figure Lengend Snippet: A maternally transmitted epigenetic factor increases diabetes incidence in the TCR+HEL+ offspring of formerly diabetic mothers. A : Diabetes incidence in TCR+HEL+ mice bred from TCR+HEL+ mothers or fathers that had spontaneously developed diabetes and had euglycemia stably restored by transplanting nontransgenic islets. The diabetes-cured TCR+HEL+ parents were bred with nontransgenic partners. Statistical analysis used the log-rank method. B : Diabetes incidence for the cohort described in ( A ) stratified by sex of the offspring. C : For each of 20 diabetes-cured TCR+HEL+ female breeders used to generate the cohort described in ( A ), two serial serum anti-HEL IgG titers (connected by a line) are plotted against the time of blood sampling relative to islet transplantation on day 0 ( left ). Serum anti-HEL IgG titers for 20 nondiabetic TCR+HEL+ females from the same colony as the diabetes-cured females ( right ).

    Article Snippet: Serial serum dilutions were applied followed by alkaline phosphatase-conjugated goat anti-mouse IgG or IgG2a (Southern Biotechnology) and then Phosphatase Substrate (Sigma).

    Techniques: Mouse Assay, Stable Transfection, Sampling, Transplantation Assay