hrp conjugated goat anti mouse igg  (Proteintech)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Goat anti mouse IgG H L HRP conjugate
    Description:

    Catalog Number:
    SA00001-1
    Price:
    50
    Applications:
    ELISA, WB, IHC/ICC
    Purity:
    The antibody was purified from antisera by immunoaffinity chromatography using antigens coupled to agarose beads.
    Conjugate:
    Purified from horseradish root
    Size:
    200ul
    Buy from Supplier


    Structured Review

    Proteintech hrp conjugated goat anti mouse igg
    “Self-cleaving” 2A peptide cleaves EYFP and RFP in EtER. A EtER sporozoites were reacted with mouse anti-SAG13 polyclonal antibody, followed by the reaction with AMCA-conjugated goat anti-mouse <t>IgG</t> (H + L) (Proteintech, USA), revealed by a confocal laser scanning microscopy (SP5, Leica, Germany). EYFP mainly localized to the nucleus, while RFP was found in the cytoplasm. Bar 5 μm. B RFP was secreted into PV in trophozoites (24 hpi) and 1 st -genernation schizont stages, while EYFP in the nucleus (48 and 72 hpi). Bar 5 μm. C Soluble proteins extracted from EtER and the wild-type E. tenella (WT) were resolved by SDS-PAGE and the immunoblot analysis was conducted following standard protocols. The primary antibody was the mouse anti-EYFP polyclonal antibody and mouse anti-His tag monoclonal antibody, the mouse anti-GAPDH polyclonal antibody served as the loading control, while the <t>HRP-conjugated</t> goat anti-mouse IgG was used as the secondary antibody. TgDHFR-EYFP was 95 kDa and RFP 28 kDa. M: marker.

    https://www.bioz.com/result/hrp conjugated goat anti mouse igg/product/Proteintech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated goat anti mouse igg - by Bioz Stars, 2021-06
    99/100 stars

    Images

    1) Product Images from "“Self-cleaving” 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella"

    Article Title: “Self-cleaving” 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella

    Journal: Veterinary Research

    doi: 10.1186/s13567-016-0351-z

    “Self-cleaving” 2A peptide cleaves EYFP and RFP in EtER. A EtER sporozoites were reacted with mouse anti-SAG13 polyclonal antibody, followed by the reaction with AMCA-conjugated goat anti-mouse IgG (H + L) (Proteintech, USA), revealed by a confocal laser scanning microscopy (SP5, Leica, Germany). EYFP mainly localized to the nucleus, while RFP was found in the cytoplasm. Bar 5 μm. B RFP was secreted into PV in trophozoites (24 hpi) and 1 st -genernation schizont stages, while EYFP in the nucleus (48 and 72 hpi). Bar 5 μm. C Soluble proteins extracted from EtER and the wild-type E. tenella (WT) were resolved by SDS-PAGE and the immunoblot analysis was conducted following standard protocols. The primary antibody was the mouse anti-EYFP polyclonal antibody and mouse anti-His tag monoclonal antibody, the mouse anti-GAPDH polyclonal antibody served as the loading control, while the HRP-conjugated goat anti-mouse IgG was used as the secondary antibody. TgDHFR-EYFP was 95 kDa and RFP 28 kDa. M: marker.
    Figure Legend Snippet: “Self-cleaving” 2A peptide cleaves EYFP and RFP in EtER. A EtER sporozoites were reacted with mouse anti-SAG13 polyclonal antibody, followed by the reaction with AMCA-conjugated goat anti-mouse IgG (H + L) (Proteintech, USA), revealed by a confocal laser scanning microscopy (SP5, Leica, Germany). EYFP mainly localized to the nucleus, while RFP was found in the cytoplasm. Bar 5 μm. B RFP was secreted into PV in trophozoites (24 hpi) and 1 st -genernation schizont stages, while EYFP in the nucleus (48 and 72 hpi). Bar 5 μm. C Soluble proteins extracted from EtER and the wild-type E. tenella (WT) were resolved by SDS-PAGE and the immunoblot analysis was conducted following standard protocols. The primary antibody was the mouse anti-EYFP polyclonal antibody and mouse anti-His tag monoclonal antibody, the mouse anti-GAPDH polyclonal antibody served as the loading control, while the HRP-conjugated goat anti-mouse IgG was used as the secondary antibody. TgDHFR-EYFP was 95 kDa and RFP 28 kDa. M: marker.

    Techniques Used: Confocal Laser Scanning Microscopy, SDS Page, Marker

    2) Product Images from "Immunogenicity and protective efficacy of DMT liposome-adjuvanted tuberculosis subunit CTT3H vaccine"

    Article Title: Immunogenicity and protective efficacy of DMT liposome-adjuvanted tuberculosis subunit CTT3H vaccine

    Journal: Human Vaccines & Immunotherapeutics

    doi: 10.1080/21645515.2015.1037057

    The levels of serum IgG, IgG1, and IgG2a antibodies against CTT3H in different groups ( n = 3). Three weeks after the last immunization, anti-CTT3H IgG, IgG1, and IgG2a (replaced by IgG2c) antibody titers in individual samples from different vaccinated
    Figure Legend Snippet: The levels of serum IgG, IgG1, and IgG2a antibodies against CTT3H in different groups ( n = 3). Three weeks after the last immunization, anti-CTT3H IgG, IgG1, and IgG2a (replaced by IgG2c) antibody titers in individual samples from different vaccinated

    Techniques Used:

    3) Product Images from "Secreted phospholipase A2 of Clonorchis sinensis activates hepatic stellate cells through a pathway involving JNK signalling"

    Article Title: Secreted phospholipase A2 of Clonorchis sinensis activates hepatic stellate cells through a pathway involving JNK signalling

    Journal: Parasites & Vectors

    doi: 10.1186/s13071-017-2082-z

    Collagen I produced by HSCs are increased after incubated with MBP- Cs sPLA2. The effect of MBP- Cs sPLA2 on activation of hepatic stellate cells and production of collagen I was evaluated by ELISA. After 36 h or 48 h, the supernatant of cells was detected by ELISA. Collagen I rabbit antibody (1:2000 dilutions) was used as the first antibody and HRP-conjugated goat anti-rabbit IgG was used as a secondary antibody (1:10,000 dilution). a Supernatant of cell culture after incubation for 36 h; b supernatant of cell culture after incubation for 48 h. Unpaired t -test was applied for statistical analysis
    Figure Legend Snippet: Collagen I produced by HSCs are increased after incubated with MBP- Cs sPLA2. The effect of MBP- Cs sPLA2 on activation of hepatic stellate cells and production of collagen I was evaluated by ELISA. After 36 h or 48 h, the supernatant of cells was detected by ELISA. Collagen I rabbit antibody (1:2000 dilutions) was used as the first antibody and HRP-conjugated goat anti-rabbit IgG was used as a secondary antibody (1:10,000 dilution). a Supernatant of cell culture after incubation for 36 h; b supernatant of cell culture after incubation for 48 h. Unpaired t -test was applied for statistical analysis

    Techniques Used: Produced, Incubation, Activation Assay, Enzyme-linked Immunosorbent Assay, Cell Culture

    Identification of protein MBP- Cs sPLA2 by western blot and mass spectrum. a The recombinant protein MBP- Cs sPLA2 was identified by western blot with Cs sPLA2 monoclonal antibody. Cs sPLA2 monoclonal antibody (1:3,000 dilution) was used as first antibody and goat anti-mouse immunoglobulin G conjugated HRP (1:2000 dilution) was used as the second antibody. Lane 1: the recombinant protein MBP- Cs sPLA2; Lane 2: protein MBP. b The recombinant protein MBP- Cs sPLA2 was identified by mass spectrometry
    Figure Legend Snippet: Identification of protein MBP- Cs sPLA2 by western blot and mass spectrum. a The recombinant protein MBP- Cs sPLA2 was identified by western blot with Cs sPLA2 monoclonal antibody. Cs sPLA2 monoclonal antibody (1:3,000 dilution) was used as first antibody and goat anti-mouse immunoglobulin G conjugated HRP (1:2000 dilution) was used as the second antibody. Lane 1: the recombinant protein MBP- Cs sPLA2; Lane 2: protein MBP. b The recombinant protein MBP- Cs sPLA2 was identified by mass spectrometry

    Techniques Used: Western Blot, Recombinant, Mass Spectrometry

    Related Articles

    SDS Page:

    Article Title: “Self-cleaving” 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella
    Article Snippet: To further test the self-cleavage efficiency of P2A in EtER, we used Western blotting. .. EtER sporozoite soluble antigens were subjected to SDS-PAGE, followed by reactions with mouse anti-EYFP polyclonal-antibody, mouse anti-His tag monoclonal antibody and HRP-conjugated goat anti-mouse IgG (Proteintech, USA) secondary antibody. ..

    Western Blot:

    Article Title: Immunogenicity and protective efficacy of DMT liposome-adjuvanted tuberculosis subunit CTT3H vaccine
    Article Snippet: The recombinant plasmid pET30b-CTT3H was transformed into E. coli strain BL21 (DE3), and the expression of CTT3H was induced by isopropylthio-β-galactoside (IPTG) at a final concentration of 1 mM for 4 h. Recombinant CTT3H protein was purified using nitrilotriacetic acid (NTA) metal ion affinity chromatography according to the manufacturer's instructions (GE Healthcare, Cat. no. 17-5318-02) and verified by 15% SDS-PAGE. .. Western blotting was used to confirm the specificity of purified proteins using anti-His 6 mouse mAb (diluted 1/2000; Tiangen, Cat. no. AB102) or mouse anti-CTT3H sera as the primary antibody and peroxidase-conjugated goat anti-mouse IgG (diluted 1/5000; Proteintech Biotech, Cat. no. SA00001-1) as the secondary antibody, respectively. .. The immunoblots were visualized using BeyoECL Plus (Beyotime, Cat. no. P0018).

    Purification:

    Article Title: Immunogenicity and protective efficacy of DMT liposome-adjuvanted tuberculosis subunit CTT3H vaccine
    Article Snippet: The recombinant plasmid pET30b-CTT3H was transformed into E. coli strain BL21 (DE3), and the expression of CTT3H was induced by isopropylthio-β-galactoside (IPTG) at a final concentration of 1 mM for 4 h. Recombinant CTT3H protein was purified using nitrilotriacetic acid (NTA) metal ion affinity chromatography according to the manufacturer's instructions (GE Healthcare, Cat. no. 17-5318-02) and verified by 15% SDS-PAGE. .. Western blotting was used to confirm the specificity of purified proteins using anti-His 6 mouse mAb (diluted 1/2000; Tiangen, Cat. no. AB102) or mouse anti-CTT3H sera as the primary antibody and peroxidase-conjugated goat anti-mouse IgG (diluted 1/5000; Proteintech Biotech, Cat. no. SA00001-1) as the secondary antibody, respectively. .. The immunoblots were visualized using BeyoECL Plus (Beyotime, Cat. no. P0018).

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein
    Article Snippet: The protein concentration was determined by the Bradford method with bovine serum albumin (BSA) as the standard. .. Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China). .. The reactions were visualized using TMB reagent (Beyotime, Nantong, China), stopped by addition of 2 M H2 SO4 , and the absorbance was measured at 450 nm using an ELx808 absorbance reader (BioTek, Winooski, VT, USA).

    Article Title: Soluble expression and purification of Bluetongue Virus Type 1 (BTV1) structure protein VP2 in Escherichia coli and its immunogenicity in mice
    Article Snippet: The microplate was coated with 50 µl purified recombinant protein VP2 (0.05 mg/well) and primary antibodies was a series of gradient diluted serum beginning with 100-fold serum as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. .. Moreover, the microplate was coated with 50 µl purified recombinant protein VP2 (0.05 mg/well) and a series of serum collected from the first week to the eighth week after immunization, as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. .. The result of indirect ELISA was read using a Microplate Reader (BioTek Instruments, Inc, Winooski, VT, US) at 490 nm.

    Binding Assay:

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein
    Article Snippet: The protein concentration was determined by the Bradford method with bovine serum albumin (BSA) as the standard. .. Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China). .. The reactions were visualized using TMB reagent (Beyotime, Nantong, China), stopped by addition of 2 M H2 SO4 , and the absorbance was measured at 450 nm using an ELx808 absorbance reader (BioTek, Winooski, VT, USA).

    Enzyme-linked Immunosorbent Assay:

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein
    Article Snippet: The protein concentration was determined by the Bradford method with bovine serum albumin (BSA) as the standard. .. Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China). .. The reactions were visualized using TMB reagent (Beyotime, Nantong, China), stopped by addition of 2 M H2 SO4 , and the absorbance was measured at 450 nm using an ELx808 absorbance reader (BioTek, Winooski, VT, USA).

    Incubation:

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein
    Article Snippet: The protein concentration was determined by the Bradford method with bovine serum albumin (BSA) as the standard. .. Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China). .. The reactions were visualized using TMB reagent (Beyotime, Nantong, China), stopped by addition of 2 M H2 SO4 , and the absorbance was measured at 450 nm using an ELx808 absorbance reader (BioTek, Winooski, VT, USA).

    Labeling:

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein
    Article Snippet: The protein concentration was determined by the Bradford method with bovine serum albumin (BSA) as the standard. .. Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China). .. The reactions were visualized using TMB reagent (Beyotime, Nantong, China), stopped by addition of 2 M H2 SO4 , and the absorbance was measured at 450 nm using an ELx808 absorbance reader (BioTek, Winooski, VT, USA).

    Recombinant:

    Article Title: Soluble expression and purification of Bluetongue Virus Type 1 (BTV1) structure protein VP2 in Escherichia coli and its immunogenicity in mice
    Article Snippet: The microplate was coated with 50 µl purified recombinant protein VP2 (0.05 mg/well) and primary antibodies was a series of gradient diluted serum beginning with 100-fold serum as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. .. Moreover, the microplate was coated with 50 µl purified recombinant protein VP2 (0.05 mg/well) and a series of serum collected from the first week to the eighth week after immunization, as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. .. The result of indirect ELISA was read using a Microplate Reader (BioTek Instruments, Inc, Winooski, VT, US) at 490 nm.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Proteintech hrp conjugated goat anti mouse igg
    “Self-cleaving” 2A peptide cleaves EYFP and RFP in EtER. A EtER sporozoites were reacted with mouse anti-SAG13 polyclonal antibody, followed by the reaction with AMCA-conjugated goat anti-mouse <t>IgG</t> (H + L) (Proteintech, USA), revealed by a confocal laser scanning microscopy (SP5, Leica, Germany). EYFP mainly localized to the nucleus, while RFP was found in the cytoplasm. Bar 5 μm. B RFP was secreted into PV in trophozoites (24 hpi) and 1 st -genernation schizont stages, while EYFP in the nucleus (48 and 72 hpi). Bar 5 μm. C Soluble proteins extracted from EtER and the wild-type E. tenella (WT) were resolved by SDS-PAGE and the immunoblot analysis was conducted following standard protocols. The primary antibody was the mouse anti-EYFP polyclonal antibody and mouse anti-His tag monoclonal antibody, the mouse anti-GAPDH polyclonal antibody served as the loading control, while the <t>HRP-conjugated</t> goat anti-mouse IgG was used as the secondary antibody. TgDHFR-EYFP was 95 kDa and RFP 28 kDa. M: marker.
    Hrp Conjugated Goat Anti Mouse Igg, supplied by Proteintech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hrp conjugated goat anti mouse igg/product/Proteintech
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hrp conjugated goat anti mouse igg - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    Image Search Results


    “Self-cleaving” 2A peptide cleaves EYFP and RFP in EtER. A EtER sporozoites were reacted with mouse anti-SAG13 polyclonal antibody, followed by the reaction with AMCA-conjugated goat anti-mouse IgG (H + L) (Proteintech, USA), revealed by a confocal laser scanning microscopy (SP5, Leica, Germany). EYFP mainly localized to the nucleus, while RFP was found in the cytoplasm. Bar 5 μm. B RFP was secreted into PV in trophozoites (24 hpi) and 1 st -genernation schizont stages, while EYFP in the nucleus (48 and 72 hpi). Bar 5 μm. C Soluble proteins extracted from EtER and the wild-type E. tenella (WT) were resolved by SDS-PAGE and the immunoblot analysis was conducted following standard protocols. The primary antibody was the mouse anti-EYFP polyclonal antibody and mouse anti-His tag monoclonal antibody, the mouse anti-GAPDH polyclonal antibody served as the loading control, while the HRP-conjugated goat anti-mouse IgG was used as the secondary antibody. TgDHFR-EYFP was 95 kDa and RFP 28 kDa. M: marker.

    Journal: Veterinary Research

    Article Title: “Self-cleaving” 2A peptide from porcine teschovirus-1 mediates cleavage of dual fluorescent proteins in transgenic Eimeria tenella

    doi: 10.1186/s13567-016-0351-z

    Figure Lengend Snippet: “Self-cleaving” 2A peptide cleaves EYFP and RFP in EtER. A EtER sporozoites were reacted with mouse anti-SAG13 polyclonal antibody, followed by the reaction with AMCA-conjugated goat anti-mouse IgG (H + L) (Proteintech, USA), revealed by a confocal laser scanning microscopy (SP5, Leica, Germany). EYFP mainly localized to the nucleus, while RFP was found in the cytoplasm. Bar 5 μm. B RFP was secreted into PV in trophozoites (24 hpi) and 1 st -genernation schizont stages, while EYFP in the nucleus (48 and 72 hpi). Bar 5 μm. C Soluble proteins extracted from EtER and the wild-type E. tenella (WT) were resolved by SDS-PAGE and the immunoblot analysis was conducted following standard protocols. The primary antibody was the mouse anti-EYFP polyclonal antibody and mouse anti-His tag monoclonal antibody, the mouse anti-GAPDH polyclonal antibody served as the loading control, while the HRP-conjugated goat anti-mouse IgG was used as the secondary antibody. TgDHFR-EYFP was 95 kDa and RFP 28 kDa. M: marker.

    Article Snippet: EtER sporozoite soluble antigens were subjected to SDS-PAGE, followed by reactions with mouse anti-EYFP polyclonal-antibody, mouse anti-His tag monoclonal antibody and HRP-conjugated goat anti-mouse IgG (Proteintech, USA) secondary antibody.

    Techniques: Confocal Laser Scanning Microscopy, SDS Page, Marker

    Molecular design and characterization of ferritin NP vaccine. a , Schematic illustration of ferritin-based vaccine delivery vehicle. b , Size exclusion chromatography of SpyTag–ferritin NPs. The ultraviolet absorptions at 280 nm and 260 nm are shown. AU, absorbance unit. c , SDS–polyacrylamide gel electrophoresis analysis of the covalent ligation of SpyTag–ferritin (~22.4 kDa) and SC–preS1 (~25.4 kDa) at 1:0.5, 1:1 and 1:2 mole ratios at 4 °C overnight. d , Size exclusion chromatography of ferritin NP–preS1. e , TEM images and two-dimensional reconstruction of SpyTag–ferritin NP and ferritin NP–preS1. Panels b – e show representative results of at least three independent experiments. f , Anti-preS1 response at vaccine immunization on day 21 was analysed using enzyme-linked immunosorbent assay (ELISA) ( n = 3 mice per group). Data are representative of five independent experiments. g , Kinetics of anti-preS1 levels after immunization ( n = 3). The red arrows indicate the immunization times. h , Comparison of conjugated and unconjugated vaccines on anti-preS1 response. i , Anti-preS1 IgG avidity on vaccine immunization was detected with ELISA. Each curve represents an individual serum sample ( n = 3) (left). The slopes of each fitting curve were calculated (right). OD, optical density. j , Titres of preS1-specific IgG1 and IgG2c in the sera from ferritin NP–preS1 vaccine ( n = 8) or SC–preS1 ( n = 14) immunized mice on day 21. g – j show representative results of three independent experiments. In f – j , data are shown as mean ± s.e.m.; statistical significance was determined using an unpaired two-tailed t -test. . Source data

    Journal: Nature Nanotechnology

    Article Title: Dual-targeting nanoparticle vaccine elicits a therapeutic antibody response against chronic hepatitis B

    doi: 10.1038/s41565-020-0648-y

    Figure Lengend Snippet: Molecular design and characterization of ferritin NP vaccine. a , Schematic illustration of ferritin-based vaccine delivery vehicle. b , Size exclusion chromatography of SpyTag–ferritin NPs. The ultraviolet absorptions at 280 nm and 260 nm are shown. AU, absorbance unit. c , SDS–polyacrylamide gel electrophoresis analysis of the covalent ligation of SpyTag–ferritin (~22.4 kDa) and SC–preS1 (~25.4 kDa) at 1:0.5, 1:1 and 1:2 mole ratios at 4 °C overnight. d , Size exclusion chromatography of ferritin NP–preS1. e , TEM images and two-dimensional reconstruction of SpyTag–ferritin NP and ferritin NP–preS1. Panels b – e show representative results of at least three independent experiments. f , Anti-preS1 response at vaccine immunization on day 21 was analysed using enzyme-linked immunosorbent assay (ELISA) ( n = 3 mice per group). Data are representative of five independent experiments. g , Kinetics of anti-preS1 levels after immunization ( n = 3). The red arrows indicate the immunization times. h , Comparison of conjugated and unconjugated vaccines on anti-preS1 response. i , Anti-preS1 IgG avidity on vaccine immunization was detected with ELISA. Each curve represents an individual serum sample ( n = 3) (left). The slopes of each fitting curve were calculated (right). OD, optical density. j , Titres of preS1-specific IgG1 and IgG2c in the sera from ferritin NP–preS1 vaccine ( n = 8) or SC–preS1 ( n = 14) immunized mice on day 21. g – j show representative results of three independent experiments. In f – j , data are shown as mean ± s.e.m.; statistical significance was determined using an unpaired two-tailed t -test. . Source data

    Article Snippet: Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (H + L) (1:5,000, ZSGB-BIO), goat anti-mouse IgG1 (1:5,000, ProteinTech), goat anti-mouse IgG2c (1:5,000, ProteinTech) and goat anti-mouse IgM (1:5,000, ProteinTech) were used to detect each isotype.

    Techniques: Size-exclusion Chromatography, Polyacrylamide Gel Electrophoresis, Ligation, Transmission Electron Microscopy, Enzyme-linked Immunosorbent Assay, Mouse Assay, Two Tailed Test

    The immune effect of fusion protein VP2 was analyzed by dot-ELISA and indirect ELISA assay. The nitrocellulose filter membrane was coated by recombinant protein VP2 and inactivated BTV1. The sera of mice and HRP-conjugated goat anti-mouse IgG antibody were used as the primary antibody and the secondary antibody, respectively. Dot-ELISA analysis is shown (A); the microplate was coated with recombinant protein VP2 and primary antibodies was a series of gradient diluted mouse serum (mouse serum collected after the third immunization ) as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. The serum titer with VP2 as coating protein is shown (B); the microplate was coated with 50 µl purified recombinant protein VP2 and primary antibodies was a series of serum collected from the first week to the eighth week after immunization, as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. Anti-VP2 antibody titer changes with immunization time (C).

    Journal: PeerJ

    Article Title: Soluble expression and purification of Bluetongue Virus Type 1 (BTV1) structure protein VP2 in Escherichia coli and its immunogenicity in mice

    doi: 10.7717/peerj.10543

    Figure Lengend Snippet: The immune effect of fusion protein VP2 was analyzed by dot-ELISA and indirect ELISA assay. The nitrocellulose filter membrane was coated by recombinant protein VP2 and inactivated BTV1. The sera of mice and HRP-conjugated goat anti-mouse IgG antibody were used as the primary antibody and the secondary antibody, respectively. Dot-ELISA analysis is shown (A); the microplate was coated with recombinant protein VP2 and primary antibodies was a series of gradient diluted mouse serum (mouse serum collected after the third immunization ) as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. The serum titer with VP2 as coating protein is shown (B); the microplate was coated with 50 µl purified recombinant protein VP2 and primary antibodies was a series of serum collected from the first week to the eighth week after immunization, as well as the secondary antibody was HRP-conjugated goat anti-mouse IgG antibody. Anti-VP2 antibody titer changes with immunization time (C).

    Article Snippet: The 200-fold diluted positive serum and 1000-fold diluted HRP-conjugated goat anti-mouse IgG antibody (Proteintech, US) were used as the primary antibody and the secondary antibody, respectively.

    Techniques: Enzyme-linked Immunosorbent Assay, Indirect ELISA, Recombinant, Mouse Assay, Purification

    Binding of Dendrolimus punctatus cypovirus ( DpCPV) virions to Spodoptera exigua midgut brush border membrane vesicles ( Se BBMVs). Se BBMVs (10 µg/mL) were coated on 96-well plates and incubated with the amount of purified DpCPV virions indicated. Virion binding was determined using a rabbit anti-DpCPV virion antibody followed by horseradish peroxidase (HRP)-labeled anti-mouse IgG as the secondary antibody. Nonspecific binding was determined with bovine serum albumin (BSA) coated wells instead of those coated with BBMVs.

    Journal: Viruses

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein

    doi: 10.3390/v9040066

    Figure Lengend Snippet: Binding of Dendrolimus punctatus cypovirus ( DpCPV) virions to Spodoptera exigua midgut brush border membrane vesicles ( Se BBMVs). Se BBMVs (10 µg/mL) were coated on 96-well plates and incubated with the amount of purified DpCPV virions indicated. Virion binding was determined using a rabbit anti-DpCPV virion antibody followed by horseradish peroxidase (HRP)-labeled anti-mouse IgG as the secondary antibody. Nonspecific binding was determined with bovine serum albumin (BSA) coated wells instead of those coated with BBMVs.

    Article Snippet: Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China).

    Techniques: Binding Assay, Incubation, Purification, Labeling

    Maltose binding protein (MBP)-fusion protein binding to Se BBMVs. ( A ) Expression and purification of MBP-fusion proteins. MBP (lane 1), MBP-galactosyltransferase (GTase) (lane 2), MBP-methyltransferase (MTase) (lane 3), MBP-VP4 (lane 4), and MBP-VP5 (lane 5) were separated by 12% sodium dodecyl (lauryl) sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); ( B ) Se BBMV proteins were coated on 96-well plates and incubated with the concentrations of MBP-fusion proteins indicated. The binding efficiency was determined with an anti-MBP mAb and HRP-labeled IgG as the secondary antibody. Nonspecific binding was determined with BSA coating instead of Se BBMVs; ( C ) MBP-fusion proteins (0.3 µM) were mixed with the amounts of purified DpCPV virions indicated, then added to the Se BBMVs-coated 96-well plates, and absorption was allowed for 1 h. Protein binding was determined as described above.

    Journal: Viruses

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein

    doi: 10.3390/v9040066

    Figure Lengend Snippet: Maltose binding protein (MBP)-fusion protein binding to Se BBMVs. ( A ) Expression and purification of MBP-fusion proteins. MBP (lane 1), MBP-galactosyltransferase (GTase) (lane 2), MBP-methyltransferase (MTase) (lane 3), MBP-VP4 (lane 4), and MBP-VP5 (lane 5) were separated by 12% sodium dodecyl (lauryl) sulfate-polyacrylamide gel electrophoresis (SDS-PAGE); ( B ) Se BBMV proteins were coated on 96-well plates and incubated with the concentrations of MBP-fusion proteins indicated. The binding efficiency was determined with an anti-MBP mAb and HRP-labeled IgG as the secondary antibody. Nonspecific binding was determined with BSA coating instead of Se BBMVs; ( C ) MBP-fusion proteins (0.3 µM) were mixed with the amounts of purified DpCPV virions indicated, then added to the Se BBMVs-coated 96-well plates, and absorption was allowed for 1 h. Protein binding was determined as described above.

    Article Snippet: Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China).

    Techniques: Binding Assay, Protein Binding, Expressing, Purification, Polyacrylamide Gel Electrophoresis, SDS Page, Incubation, Labeling

    The MTase binding protein was identified as B. mori alkaline phosphatase protein (ALP). ( A ) Far-Western blotting of Bm BBMVs with MBP-MTase. Blot of Bm BBMVs were separated by 10% SDS-PAGE (lane 1), incubated with MBP (lane 2) or MBP-MTase (lane 3), and probed with an anti-MBP mAb followed by HRP-labeled anti-mouse IgG as the secondary antibody; ( B ) Expression and purification of recombinant Bm ALP protein separated by 12% SDS-PAGE. Lanes: M, protein molecular mass marker; 1, non-induced bacterial cell lysate; 2, induced bacterial cell lysate; 3, Bm ALP purified by a Ni-NTA agarose column; 4, Bm ALP detected with a rabbit anti-ALP antibody; ( C ) Co-immunoprecipitation (Co-IP) validated the interaction between MBP-MTase and Bm ALP. The immunoprecipitated products (lane 1 to lane 4) and Bm ALP protein (lane 5) were separated by SDS-PAGE, transferred to a PVDF membrane, and were incubated with a mixture of mouse anti-MBP mAb and mouse anti-His mAb as the primary antibody and HRP-labeled anti-mouse IgG as the secondary antibody.

    Journal: Viruses

    Article Title: MTase Domain of Dendrolimus punctatus cypovirus VP3 Mediates Virion Attachment and Interacts with Host ALP Protein

    doi: 10.3390/v9040066

    Figure Lengend Snippet: The MTase binding protein was identified as B. mori alkaline phosphatase protein (ALP). ( A ) Far-Western blotting of Bm BBMVs with MBP-MTase. Blot of Bm BBMVs were separated by 10% SDS-PAGE (lane 1), incubated with MBP (lane 2) or MBP-MTase (lane 3), and probed with an anti-MBP mAb followed by HRP-labeled anti-mouse IgG as the secondary antibody; ( B ) Expression and purification of recombinant Bm ALP protein separated by 12% SDS-PAGE. Lanes: M, protein molecular mass marker; 1, non-induced bacterial cell lysate; 2, induced bacterial cell lysate; 3, Bm ALP purified by a Ni-NTA agarose column; 4, Bm ALP detected with a rabbit anti-ALP antibody; ( C ) Co-immunoprecipitation (Co-IP) validated the interaction between MBP-MTase and Bm ALP. The immunoprecipitated products (lane 1 to lane 4) and Bm ALP protein (lane 5) were separated by SDS-PAGE, transferred to a PVDF membrane, and were incubated with a mixture of mouse anti-MBP mAb and mouse anti-His mAb as the primary antibody and HRP-labeled anti-mouse IgG as the secondary antibody.

    Article Snippet: Binding Characteristics of S. exigua BBMVs and Competition Assays ELISA plates (96-well) were coated with 10 µg/mL of S. exigua BBMVs (Se BBMVs) in carbonate buffer at 4 °C overnight and blocked with 3% BSA for 2 h. Increasing amounts of purified DpCPV virions (0.01–100 µg) or MBP-fusion proteins (0.02–4 µM) in a total volume of 100 µL were added to the wells and incubated for 1 h. The wells were washed with PBST (4 mM KH2 PO4 , 16 mM Na2 HPO4 , 115 mM NaCl, 0.05% Tween 20, pH 7.4) and incubated with a rabbit anti-DpCPV virion antibody or a mouse anti-MBP mAb (Sigma-Aldrich, St. Louis, MO, USA) followed by horseradish peroxidase (HRP) labeled anti-rabbit IgG (Sigma-Aldrich) or HRP labeled anti-mouse IgG (Proteintech, Wuhan, China).

    Techniques: Binding Assay, ALP Assay, Far Western Blot, SDS Page, Incubation, Labeling, Expressing, Purification, Recombinant, Marker, Immunoprecipitation, Co-Immunoprecipitation Assay