Structured Review

Agilent technologies hrp conjugated secondary antibodies
Binding of recombinant <t>CD2831</t> full-length protein and subdomains to collagen. ( A ) Schematic representation of CD2831 domains organization displays the Leader Peptide at the N-terminus (orange) followed by a Short Repeat Region (yellow) in which repeated amino acids sequences are marked in bold, two collagen-binding domains (blue) and a Proline-rich region (green) recognized by the protease ZmpI/PPEP-1 that cleaves within the seven NPP amino acids sequences of the domain. The charged tail and the hydrophobic domain are present at the C-terminus (red). The protein is cleaved between the threonyl and glycil residues of the PPKTG motif and transferred to the cell-wall peptidoglycan by the Sortase B enzyme. Blue, black and grey lines indicate the length of recombinant CD2831 subdomains compared to the full-length. ( B ) Immunofluorescence of 1 µM of CD2831 (green) binding to collagen (red) produced by IMR90 cells. Collagen staining results from a mix of antibodies against collagen types I, III and V in a 1:1:1 ratio. Binding of the proteins was detected by using rabbit anti-CD2831 serum and secondary Alexa Fluor-conjugated antibodies. ( C ) Human fibroblasts IMR90 were cultured into collagen I-coated 96 well plates for three days after seeding. Cells were then incubated with serially diluted recombinant CD2831 proteins ranging from 0.08 to 10 µM. Binding of CD2831 was detected as above and quantified with a microplate fluorescence reader. ( D ) ELISA plates were coated with 10 μg/ml of purified collagens and incubated with serially diluted recombinant CD2831 proteins ranging from 15 nM to 2 µM. Binding of the proteins was detected using rabbit anti-CD2831 serum, followed by <t>HRP-conjugated-secondary</t> antibody. CWP25 protein was used as negative control. CD2831 CBD1 and CD2831 CBD2 represent the previously described subdomains 1 and 2.
Hrp Conjugated Secondary Antibodies, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 389 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hrp conjugated secondary antibodies/product/Agilent technologies
Average 94 stars, based on 389 article reviews
Price from $9.99 to $1999.99
hrp conjugated secondary antibodies - by Bioz Stars, 2020-07
94/100 stars

Images

1) Product Images from "Dual role of the colonization factor CD2831 in Clostridium difficile pathogenesis"

Article Title: Dual role of the colonization factor CD2831 in Clostridium difficile pathogenesis

Journal: Scientific Reports

doi: 10.1038/s41598-019-42000-8

Binding of recombinant CD2831 full-length protein and subdomains to collagen. ( A ) Schematic representation of CD2831 domains organization displays the Leader Peptide at the N-terminus (orange) followed by a Short Repeat Region (yellow) in which repeated amino acids sequences are marked in bold, two collagen-binding domains (blue) and a Proline-rich region (green) recognized by the protease ZmpI/PPEP-1 that cleaves within the seven NPP amino acids sequences of the domain. The charged tail and the hydrophobic domain are present at the C-terminus (red). The protein is cleaved between the threonyl and glycil residues of the PPKTG motif and transferred to the cell-wall peptidoglycan by the Sortase B enzyme. Blue, black and grey lines indicate the length of recombinant CD2831 subdomains compared to the full-length. ( B ) Immunofluorescence of 1 µM of CD2831 (green) binding to collagen (red) produced by IMR90 cells. Collagen staining results from a mix of antibodies against collagen types I, III and V in a 1:1:1 ratio. Binding of the proteins was detected by using rabbit anti-CD2831 serum and secondary Alexa Fluor-conjugated antibodies. ( C ) Human fibroblasts IMR90 were cultured into collagen I-coated 96 well plates for three days after seeding. Cells were then incubated with serially diluted recombinant CD2831 proteins ranging from 0.08 to 10 µM. Binding of CD2831 was detected as above and quantified with a microplate fluorescence reader. ( D ) ELISA plates were coated with 10 μg/ml of purified collagens and incubated with serially diluted recombinant CD2831 proteins ranging from 15 nM to 2 µM. Binding of the proteins was detected using rabbit anti-CD2831 serum, followed by HRP-conjugated-secondary antibody. CWP25 protein was used as negative control. CD2831 CBD1 and CD2831 CBD2 represent the previously described subdomains 1 and 2.
Figure Legend Snippet: Binding of recombinant CD2831 full-length protein and subdomains to collagen. ( A ) Schematic representation of CD2831 domains organization displays the Leader Peptide at the N-terminus (orange) followed by a Short Repeat Region (yellow) in which repeated amino acids sequences are marked in bold, two collagen-binding domains (blue) and a Proline-rich region (green) recognized by the protease ZmpI/PPEP-1 that cleaves within the seven NPP amino acids sequences of the domain. The charged tail and the hydrophobic domain are present at the C-terminus (red). The protein is cleaved between the threonyl and glycil residues of the PPKTG motif and transferred to the cell-wall peptidoglycan by the Sortase B enzyme. Blue, black and grey lines indicate the length of recombinant CD2831 subdomains compared to the full-length. ( B ) Immunofluorescence of 1 µM of CD2831 (green) binding to collagen (red) produced by IMR90 cells. Collagen staining results from a mix of antibodies against collagen types I, III and V in a 1:1:1 ratio. Binding of the proteins was detected by using rabbit anti-CD2831 serum and secondary Alexa Fluor-conjugated antibodies. ( C ) Human fibroblasts IMR90 were cultured into collagen I-coated 96 well plates for three days after seeding. Cells were then incubated with serially diluted recombinant CD2831 proteins ranging from 0.08 to 10 µM. Binding of CD2831 was detected as above and quantified with a microplate fluorescence reader. ( D ) ELISA plates were coated with 10 μg/ml of purified collagens and incubated with serially diluted recombinant CD2831 proteins ranging from 15 nM to 2 µM. Binding of the proteins was detected using rabbit anti-CD2831 serum, followed by HRP-conjugated-secondary antibody. CWP25 protein was used as negative control. CD2831 CBD1 and CD2831 CBD2 represent the previously described subdomains 1 and 2.

Techniques Used: Binding Assay, Recombinant, Immunofluorescence, Produced, Staining, Cell Culture, Incubation, Fluorescence, Enzyme-linked Immunosorbent Assay, Purification, Negative Control

2) Product Images from "A receptor fusion protein for the inhibition of murine oncostatin M"

Article Title: A receptor fusion protein for the inhibition of murine oncostatin M

Journal: BMC Biotechnology

doi: 10.1186/1472-6750-11-3

Complexes of mOSM-RFPs and mOSM analyzed by bn-PAGE . (A) 35 ng of mOSM were incubated with 200 ng of the respective mOSM-RFP (with the exception of i-mOSM-RFP that is less efficiently expressed and therefore 200 ng could not be achieved) in 50 μl for 30 min. Subsequently Coomassie Brilliant blue G-250 was added and the protein complexes were separated on a native gradient gel (4-16% PAA). After blotting of the proteins to a PVDF membrane mOSM-RFPs were detected using a FLAG antibody (upper panel). After stripping of the blot mOSM was detected using a mOSM antibody (middle panel). (B) bn-PAGE was performed as described in (A) with the protein amounts indicated in the figure. After blotting, detection of the proteins was performed with primary goat-anti-mOSM and mouse-anti-flag antibodies followed by secondary rabbit-anti-goat-Cy2 and donkey-anti-mouse-Cy3 antibodies. Fluorescence was detected with a fluorescence scanner. Afterwards the mOSM antibody was visualized by ECL using a matching HRP-conjugated secondary antibody.
Figure Legend Snippet: Complexes of mOSM-RFPs and mOSM analyzed by bn-PAGE . (A) 35 ng of mOSM were incubated with 200 ng of the respective mOSM-RFP (with the exception of i-mOSM-RFP that is less efficiently expressed and therefore 200 ng could not be achieved) in 50 μl for 30 min. Subsequently Coomassie Brilliant blue G-250 was added and the protein complexes were separated on a native gradient gel (4-16% PAA). After blotting of the proteins to a PVDF membrane mOSM-RFPs were detected using a FLAG antibody (upper panel). After stripping of the blot mOSM was detected using a mOSM antibody (middle panel). (B) bn-PAGE was performed as described in (A) with the protein amounts indicated in the figure. After blotting, detection of the proteins was performed with primary goat-anti-mOSM and mouse-anti-flag antibodies followed by secondary rabbit-anti-goat-Cy2 and donkey-anti-mouse-Cy3 antibodies. Fluorescence was detected with a fluorescence scanner. Afterwards the mOSM antibody was visualized by ECL using a matching HRP-conjugated secondary antibody.

Techniques Used: Polyacrylamide Gel Electrophoresis, Incubation, Stripping Membranes, Fluorescence

Related Articles

Negative Control:

Article Title: Peptide inhibition of the SETD6 methyltransferase catalytic activity
Article Snippet: .. After 3 washes with PBST, the plate was covered with 0.5 μg GST-SETD6 or GST protein (negative control) diluted in 1% BSA in PBST for 1 h. Plates were then washed and incubated with primary antibody (anti-GST, 1:4000 dilution) followed by incubation with HRP-conjugated secondary antibody (goat anti-rabbit, 1:2000 dilution) for 1 h. Finally, TMB reagent (Dako) and then 1N H2 SO4 were added, and the absorbance at 450 nm was detected using a Tecan Infinite M200 plate reader. .. Wound healing assay Transfected cells and control cells were plated separately overnight to achieve a confluent cell layer in 6-well culture plates in duplicates.

Fluorescence:

Article Title: A receptor fusion protein for the inhibition of murine oncostatin M
Article Snippet: .. Gels were analyzed by Western blotting using antibodies directed against mOSM (goat-anti-mOSM, R & D Systems, MN, USA), FLAG (mouse-anti-flag, SIGMA-Aldrich, Taufkirchen, Germany) and HRP conjugated secondary antibodies for ECL detection or rabbit-anti-goat-Cy2 and donkey-anti-mouse-Cy3 for fluorescence detection (Dako, Hamburg, Germany). .. Fluorescence was detected with a fluorescence scanner (Typhoon, Amersham).

Blocking Assay:

Article Title: DNA immunization with fusion genes encoding different regions of hepatitis C virus E2 fused to the gene for hepatitis B surface antigen elicits immune responses to both HCV and HBV
Article Snippet: .. After blocking for 1 h with 5% powdered lipid-free milk in PBST (PBS containing 0.1% Tween 20), the membrane was incubated with anti-HBs McAb H116 (kindly provided by Dr. E. Hildt, Munich, Germany) or anti-HCV E2 (450-565) polyclonal antiserum RE2-116[ ], followed by incubation with HRP conjugated secondary antibody (Dako Co., Denmark). .. Signals were detected by enhanced chemi-luminescence (ECL) blotting analysis system (Amersham-Pharmacia Co., UK).

Article Title: Dual role of the colonization factor CD2831 in Clostridium difficile pathogenesis
Article Snippet: .. After blocking with PBS containing 0.05% Tween-20 (PBS-T) and 10% (w/v) milk (Sigma, USA), proteins were detected with rabbit anti-CD2831 serum diluted 1:1000 for 1 hour at RT, followed by the HRP-conjugated secondary antibodies (Dako, Denmark) diluted 1:2000 for 1 hour at RT. ..

Incubation:

Article Title: Peptide inhibition of the SETD6 methyltransferase catalytic activity
Article Snippet: .. After 3 washes with PBST, the plate was covered with 0.5 μg GST-SETD6 or GST protein (negative control) diluted in 1% BSA in PBST for 1 h. Plates were then washed and incubated with primary antibody (anti-GST, 1:4000 dilution) followed by incubation with HRP-conjugated secondary antibody (goat anti-rabbit, 1:2000 dilution) for 1 h. Finally, TMB reagent (Dako) and then 1N H2 SO4 were added, and the absorbance at 450 nm was detected using a Tecan Infinite M200 plate reader. .. Wound healing assay Transfected cells and control cells were plated separately overnight to achieve a confluent cell layer in 6-well culture plates in duplicates.

Article Title: DNA immunization with fusion genes encoding different regions of hepatitis C virus E2 fused to the gene for hepatitis B surface antigen elicits immune responses to both HCV and HBV
Article Snippet: .. After blocking for 1 h with 5% powdered lipid-free milk in PBST (PBS containing 0.1% Tween 20), the membrane was incubated with anti-HBs McAb H116 (kindly provided by Dr. E. Hildt, Munich, Germany) or anti-HCV E2 (450-565) polyclonal antiserum RE2-116[ ], followed by incubation with HRP conjugated secondary antibody (Dako Co., Denmark). .. Signals were detected by enhanced chemi-luminescence (ECL) blotting analysis system (Amersham-Pharmacia Co., UK).

Article Title: Generation of NSE-MerCreMer Transgenic Mice with Tamoxifen Inducible Cre Activity in Neurons
Article Snippet: .. After TBS-T rinse, and an incubation in HRP-conjugated secondary antibody (1∶5000 dilution; P0399; Dako; 1 hour, room temperature), the membrane was washed in TBS-T. Signal was visualized by chemiluminescence (GE Health Amersham ECL Plus Western Blotting Detection System) and exposure to X-ray film (Kodak). ..

Article Title: Adiponectin suppresses amyloid-β oligomer (AβO)-induced inflammatory response of microglia via AdipoR1-AMPK-NF-κB signaling pathway
Article Snippet: .. Inc., USA) at 4 °C overnight, followed by incubation with HRP-conjugated secondary antibody (goat anti-rabbit, 1:200; Dako, Glostrup, Denmark) at RT for 1 h. Sections were developed by brown color staining and counterstained with hematoxylin. .. Quantification of amyloid plaques and the number of microglia around plaques Each cryosection (40 μm thick) was stained with 0.01% thioflavin-S (Sigma-Aldrich) followed by immunostaining with anti-Iba1 (Wako Chemicals).

Article Title: The Host Defense Peptide Cathelicidin Is Required for NK Cell-Mediated Suppression of Tumor Growth
Article Snippet: .. After washing and incubation with the species-specific, HRP-conjugated secondary Ab (DakoCytomation, Carpinteria, CA), immunoreactive proteins were detected by Western Lightning system (PerkinElmer, Wellesley, MA). .. Splenocytes from eight mice were FACS sorted for NK1.1+ CD3− NK cells, NK1.1+ CD3+ NKT cells, and NK1.1− CD3+ T cells.

Article Title: FGFR3IIIS: a novel soluble FGFR3 spliced variant that modulates growth is frequently expressed in tumour cells
Article Snippet: .. Membranes were incubated with a polyclonal antibody raised against a peptide to the carboxyl terminus of FGFR3 (SC-123, 1 : 200; Santa Cruz Biotechnology Inc., Santa Cruz, California, USA) in TTBS containing 1% (w v−1 ) nonfat dried milk for 2 h at RT; the signal was detected using an HRP-conjugated secondary antibody (1 : 2000; Dako, High Wycombe, UK) for 45 min at RT and enhanced chemiluminescence (ECL, Amersham Biosciences (UK) Ltd, Bucks, UK). .. A negative control blot was incubated with the secondary antibody alone, to check for nonspecific binding of the secondary antibody.

Western Blot:

Article Title: Generation of NSE-MerCreMer Transgenic Mice with Tamoxifen Inducible Cre Activity in Neurons
Article Snippet: .. After TBS-T rinse, and an incubation in HRP-conjugated secondary antibody (1∶5000 dilution; P0399; Dako; 1 hour, room temperature), the membrane was washed in TBS-T. Signal was visualized by chemiluminescence (GE Health Amersham ECL Plus Western Blotting Detection System) and exposure to X-ray film (Kodak). ..

Article Title: The Host Defense Peptide Cathelicidin Is Required for NK Cell-Mediated Suppression of Tumor Growth
Article Snippet: .. After washing and incubation with the species-specific, HRP-conjugated secondary Ab (DakoCytomation, Carpinteria, CA), immunoreactive proteins were detected by Western Lightning system (PerkinElmer, Wellesley, MA). .. Splenocytes from eight mice were FACS sorted for NK1.1+ CD3− NK cells, NK1.1+ CD3+ NKT cells, and NK1.1− CD3+ T cells.

Article Title: A receptor fusion protein for the inhibition of murine oncostatin M
Article Snippet: .. Gels were analyzed by Western blotting using antibodies directed against mOSM (goat-anti-mOSM, R & D Systems, MN, USA), FLAG (mouse-anti-flag, SIGMA-Aldrich, Taufkirchen, Germany) and HRP conjugated secondary antibodies for ECL detection or rabbit-anti-goat-Cy2 and donkey-anti-mouse-Cy3 for fluorescence detection (Dako, Hamburg, Germany). .. Fluorescence was detected with a fluorescence scanner (Typhoon, Amersham).

Staining:

Article Title: Adiponectin suppresses amyloid-β oligomer (AβO)-induced inflammatory response of microglia via AdipoR1-AMPK-NF-κB signaling pathway
Article Snippet: .. Inc., USA) at 4 °C overnight, followed by incubation with HRP-conjugated secondary antibody (goat anti-rabbit, 1:200; Dako, Glostrup, Denmark) at RT for 1 h. Sections were developed by brown color staining and counterstained with hematoxylin. .. Quantification of amyloid plaques and the number of microglia around plaques Each cryosection (40 μm thick) was stained with 0.01% thioflavin-S (Sigma-Aldrich) followed by immunostaining with anti-Iba1 (Wako Chemicals).

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    Agilent technologies envision plus polymer anti mouse conjugated to hrp
    Representative biopsies of pediatric duodenum immune-stained for αS . Upper panels: The three top panels are serial sections (20X), stained with H E, immuno-stained for αS, or for CD68. The specimen exhibits severe acute and chronic inflammation. Lower panels: αS co-localizes with PGP 9.5 by double immunofluorescence demonstrating expression within neurons. is shown. Primary antibodies: α-syn: ab27766 from Abcam; PGP 9.5: Z5116 from DAKO. Secondary antibodies: α-syn: Envision anti-mouse <t>HRP</t> polymer from <t>Agilent;</t> PGP 9.5: Goat anti-rabbit biotin conjugated from Vector. Tertiary (fluorescent tag): α-syn: Perkin Elmer Tyramide Signal Amplification with Cy5; PGP 9.5: Life Technologies streptavidin conjugated with AlexaFluor488. Procedures were as specified by the protocols provided with the reagents. Cy5 was imaged at 555nm, GFP at 488nm. Images were taken using Slidebook 6.0 and the overlay was an output from the Slidebook System.
    Envision Plus Polymer Anti Mouse Conjugated To Hrp, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/envision plus polymer anti mouse conjugated to hrp/product/Agilent technologies
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    envision plus polymer anti mouse conjugated to hrp - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

    91
    Agilent technologies appropriate hrp conjugated secondary antibodies
    Representative biopsies of pediatric duodenum immune-stained for αS . Upper panels: The three top panels are serial sections (20X), stained with H E, immuno-stained for αS, or for CD68. The specimen exhibits severe acute and chronic inflammation. Lower panels: αS co-localizes with PGP 9.5 by double immunofluorescence demonstrating expression within neurons. is shown. Primary antibodies: α-syn: ab27766 from Abcam; PGP 9.5: Z5116 from DAKO. Secondary antibodies: α-syn: Envision anti-mouse <t>HRP</t> polymer from <t>Agilent;</t> PGP 9.5: Goat anti-rabbit biotin conjugated from Vector. Tertiary (fluorescent tag): α-syn: Perkin Elmer Tyramide Signal Amplification with Cy5; PGP 9.5: Life Technologies streptavidin conjugated with AlexaFluor488. Procedures were as specified by the protocols provided with the reagents. Cy5 was imaged at 555nm, GFP at 488nm. Images were taken using Slidebook 6.0 and the overlay was an output from the Slidebook System.
    Appropriate Hrp Conjugated Secondary Antibodies, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 91/100, based on 49 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/appropriate hrp conjugated secondary antibodies/product/Agilent technologies
    Average 91 stars, based on 49 article reviews
    Price from $9.99 to $1999.99
    appropriate hrp conjugated secondary antibodies - by Bioz Stars, 2020-07
    91/100 stars
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    94
    Agilent technologies horseradish peroxidase
    Representative biopsies of pediatric duodenum immune-stained for αS . Upper panels: The three top panels are serial sections (20X), stained with H E, immuno-stained for αS, or for CD68. The specimen exhibits severe acute and chronic inflammation. Lower panels: αS co-localizes with PGP 9.5 by double immunofluorescence demonstrating expression within neurons. is shown. Primary antibodies: α-syn: ab27766 from Abcam; PGP 9.5: Z5116 from DAKO. Secondary antibodies: α-syn: Envision anti-mouse <t>HRP</t> polymer from <t>Agilent;</t> PGP 9.5: Goat anti-rabbit biotin conjugated from Vector. Tertiary (fluorescent tag): α-syn: Perkin Elmer Tyramide Signal Amplification with Cy5; PGP 9.5: Life Technologies streptavidin conjugated with AlexaFluor488. Procedures were as specified by the protocols provided with the reagents. Cy5 was imaged at 555nm, GFP at 488nm. Images were taken using Slidebook 6.0 and the overlay was an output from the Slidebook System.
    Horseradish Peroxidase, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 94/100, based on 510 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/horseradish peroxidase/product/Agilent technologies
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    Representative biopsies of pediatric duodenum immune-stained for αS . Upper panels: The three top panels are serial sections (20X), stained with H E, immuno-stained for αS, or for CD68. The specimen exhibits severe acute and chronic inflammation. Lower panels: αS co-localizes with PGP 9.5 by double immunofluorescence demonstrating expression within neurons. is shown. Primary antibodies: α-syn: ab27766 from Abcam; PGP 9.5: Z5116 from DAKO. Secondary antibodies: α-syn: Envision anti-mouse HRP polymer from Agilent; PGP 9.5: Goat anti-rabbit biotin conjugated from Vector. Tertiary (fluorescent tag): α-syn: Perkin Elmer Tyramide Signal Amplification with Cy5; PGP 9.5: Life Technologies streptavidin conjugated with AlexaFluor488. Procedures were as specified by the protocols provided with the reagents. Cy5 was imaged at 555nm, GFP at 488nm. Images were taken using Slidebook 6.0 and the overlay was an output from the Slidebook System.

    Journal: Journal of innate immunity

    Article Title: A role for neuronal alpha synuclein in gastrointestinal immunity

    doi: 10.1159/000477990

    Figure Lengend Snippet: Representative biopsies of pediatric duodenum immune-stained for αS . Upper panels: The three top panels are serial sections (20X), stained with H E, immuno-stained for αS, or for CD68. The specimen exhibits severe acute and chronic inflammation. Lower panels: αS co-localizes with PGP 9.5 by double immunofluorescence demonstrating expression within neurons. is shown. Primary antibodies: α-syn: ab27766 from Abcam; PGP 9.5: Z5116 from DAKO. Secondary antibodies: α-syn: Envision anti-mouse HRP polymer from Agilent; PGP 9.5: Goat anti-rabbit biotin conjugated from Vector. Tertiary (fluorescent tag): α-syn: Perkin Elmer Tyramide Signal Amplification with Cy5; PGP 9.5: Life Technologies streptavidin conjugated with AlexaFluor488. Procedures were as specified by the protocols provided with the reagents. Cy5 was imaged at 555nm, GFP at 488nm. Images were taken using Slidebook 6.0 and the overlay was an output from the Slidebook System.

    Article Snippet: The secondary antibody was Envision Plus Polymer anti-mouse conjugated to HRP (Agilent K4001).

    Techniques: Staining, Immunofluorescence, Expressing, Plasmid Preparation, Amplification