biotinylated anti rabbit antibody  (Vector Laboratories)


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    Name:
    Biotinylated Rabbit Anti Rat IgG Antibody
    Description:
    Biotinylated Rabbit Anti Rat IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Rabbit Anti Rat IgG H L is supplied in lyophilized form and can be reconstituted with 1 ml water With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains This antibody is included in the VECTASTAIN ABC kits
    Catalog Number:
    BA-4000
    Price:
    None
    Category:
    Antibodies
    Reactivity:
    Rat
    Size:
    1 5 mg
    Host:
    Rabbit
    Buy from Supplier


    Structured Review

    Vector Laboratories biotinylated anti rabbit antibody
    Biotinylated Rabbit Anti Rat IgG Antibody
    Biotinylated Rabbit Anti Rat IgG Antibody is prepared using proprietary immunization schedules that produce high affinity antibodies The antibodies are then purified by affinity chromatography and cross reactivities that are likely to interfere with specific labeling are removed by solid phase adsorption techniques The biotinylated secondary antibodies are conjugated to ensure the maximum degree of labeling without compromising the specificity or affinity of the antibody These antibodies are subjected to rigorous quality control assays and can be used for tissue and cell staining ELISAs and blots Biotinylated Rabbit Anti Rat IgG H L is supplied in lyophilized form and can be reconstituted with 1 ml water With some exceptions the recommended dilution for most applications is 1 200 H L indicates the antibody recognizes both heavy and light chains This antibody is included in the VECTASTAIN ABC kits
    https://www.bioz.com/result/biotinylated anti rabbit antibody/product/Vector Laboratories
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated anti rabbit antibody - by Bioz Stars, 2021-04
    97/100 stars

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    Related Articles

    Immunofluorescence:

    Article Title: Abnormalities of Thymic Stroma may Contribute to Immune Dysregulation in Murine Models of Leaky Severe Combined Immunodeficiency
    Article Snippet: After washes, single immunostains were revealed using the ChemMATE Envision Rabbit/Mouse (DAKO Cytomation, Glostrup, Denmark) or NovoLinkTM Polymer Detection System (NovocastraTM Laboratories Ltd, Newcastle Upon Tyne, United Kingdom) followed by diaminobenzidine (DAB) as chromogen and Hematoxylin as counterstain. .. The same procedure was applied to double immunofluorescence stainings prior to the incubation with a secondary swine anti-rabbit FITC-conjugated antibody (1:30; DAKO Cytomation, Glostrup, Denmark) for CK5 and a rabbit anti-rat biotinylated antibody (1:200; Vector Laboratories, Burlingame, CA, USA) followed by Streptavidin–Texas Red (1:100; Southern Biotechnology Associates, Birmingham, AL, USA) for CK8. ..

    Article Title: ID family protein expression and regulation in hypoxic pulmonary hypertension
    Article Snippet: .. Immunofluorescence was performed on formalin-fixed, paraffin-embedded lung sections with the ID1–3 antibodies outlined above and detected with a rabbit IgG-specific biotin-conjugated secondary antibody, followed by streptavidin-conjugated HRP (ABC kit, Vector Laboratories). .. Cy3-labeled tyramide signal amplification was used to amplify the HRP signal (Perkin Elmer, NEL744), according to the manufacturers' instructions.

    Incubation:

    Article Title: Abnormalities of Thymic Stroma may Contribute to Immune Dysregulation in Murine Models of Leaky Severe Combined Immunodeficiency
    Article Snippet: After washes, single immunostains were revealed using the ChemMATE Envision Rabbit/Mouse (DAKO Cytomation, Glostrup, Denmark) or NovoLinkTM Polymer Detection System (NovocastraTM Laboratories Ltd, Newcastle Upon Tyne, United Kingdom) followed by diaminobenzidine (DAB) as chromogen and Hematoxylin as counterstain. .. The same procedure was applied to double immunofluorescence stainings prior to the incubation with a secondary swine anti-rabbit FITC-conjugated antibody (1:30; DAKO Cytomation, Glostrup, Denmark) for CK5 and a rabbit anti-rat biotinylated antibody (1:200; Vector Laboratories, Burlingame, CA, USA) followed by Streptavidin–Texas Red (1:100; Southern Biotechnology Associates, Birmingham, AL, USA) for CK8. ..

    Article Title: Macrophage lipoprotein lipase modulates the development of atherosclerosis but not adiposity
    Article Snippet: Cross-sections of aortic roots were incubated with primary antibodies for mouse MOMA-2 (1:600; Accurate Chemical and Scientific) overnight at 4°C to investigate macrophage infiltration in the atherosclerotic lesion. .. After washing, sections were incubated with biotinylated anti-rat antibody for 1 h at room temperature, and then with avidin-biotin peroxidase complex (Vector Labs) for 30 min. Lastly, sections were developed with 3,3′-diaminobenzidine tetrahydrochloride (DAB) (Sigma), and counterstained with hematoxylin. .. The adipose tissue of mice fed a WTD for 12 weeks was fixed with 10% neutral-buffered formalin, embedded in paraffin, and sections were stained with hematoxylin and eosin.

    Article Title: Immunohistochemical analysis of macroautophagy
    Article Snippet: Subsequently, the primary antibody was applied overnight in a damp box, followed by 2 × 5 min immersion in staining buffer. .. Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer. .. In case the primary antibody was already biotinylated, blocking with normal serum and incubation with biotinylated secondary antibodies were omitted.

    Article Title: Melatonin MT1 and MT2 Receptors in the Ram Reproductive Tract
    Article Snippet: The sections were then washed in PBS pH 7.4 and incubated in normal horse serum as a blocking reagent for 40 min, followed by overnight incubation at 4 °C with the specific primary antibody (rabbit polyclonal antibody against the MT1 receptor (RRID:AB_1241048, GeneTex, Irvine, CA, USA) and rabbit polyclonal antibody against the MT2 receptor (RRID:AB_1619198, Acris Antibodies GmbH, Herford, Germany), both at a dilution of 1:50 in PBS. .. Subsequently, the slides were incubated with biotinylated anti-rabbit antiserum (Vector Laboratories) for 30 min. An avidin-biotin-peroxidase complex (Vector Laboratories) was then applied for 40 min. .. The binding sites of the primary antibodies were visualized by diaminobenzidine (DAB) and peroxidase solution (0.12 g DAB in 240 mL of PBS pH 7.4 containing 3% H2 O2 ) for 7 min.

    Article Title: Knockdown of L Calcium Channel Subtypes: Differential Effects in Neuropathic Pain
    Article Snippet: .. They were then incubated with a biotinylated anti-rabbit or anti-mouse secondary antibody (Vector Laboratories) (1:100 in PBS-NaI-BSA) for 30 min at room temperature. .. Sections were rinsed in PBS-NaI-BSA and then incubated with radiolabeled (35 S) streptavidin (GE Healthcare Biosciences) (1:100 in PBS-NaI-BSA) for 30 min at room temperature.

    Formalin-fixed Paraffin-Embedded:

    Article Title: ID family protein expression and regulation in hypoxic pulmonary hypertension
    Article Snippet: .. Immunofluorescence was performed on formalin-fixed, paraffin-embedded lung sections with the ID1–3 antibodies outlined above and detected with a rabbit IgG-specific biotin-conjugated secondary antibody, followed by streptavidin-conjugated HRP (ABC kit, Vector Laboratories). .. Cy3-labeled tyramide signal amplification was used to amplify the HRP signal (Perkin Elmer, NEL744), according to the manufacturers' instructions.

    Labeling:

    Article Title: Endocytosis of a Glycosylphosphatidylinositol-anchored Protein via Clathrin-coated Vesicles, Sorting by Default in Endosomes, and Exocytosis via RAB11-positive Carriers V⃞
    Article Snippet: For double labeling (Figures , inset; ), the first antibody was generally detected by PAG-10 or PAG-6; after a block with a rabbit normal IgG (1:400), the second antibody was detected by PAG-10 or PAG-15. .. In one case ( , inset), clathrin was first labeled with rabbit anti-TbCLH antibodies and PAG-6, whereas VSG was subsequently detected using biotinylated rabbit anti-VSG IgG followed by a monoclonal mouse antibiotin antibody (1:100; Vector Laboratories, Burlingame, CA) and goat anti-mouse IgG-18-nm gold conjugates (1:20; Jackson Immunoresearch Laboratories, West Grove, PA). .. After labeling, the cryosections were embedded in 1.4% methylcellulose, 0.3% uranyl acetate.

    Avidin-Biotin Assay:

    Article Title: Anti-α4β1 Integrin Antibodies Attenuated Brain Inflammatory Changes in a Mouse Model of Alzheimer’s Disease
    Article Snippet: All mouse cytokine ELISA kits were obtained from R & D Systems (Minneapolis, MN). .. Elite Vectastain ABC avidin and biotin kits, biotinylated anti-rabbit, anti-mouse, and anti-rat antibodies and the Vector VIP kits were from Vector Laboratories Inc. (Burlingame, CA). .. Anti-CD68 was obtained from Serotec (Raleigh, NC).

    Article Title: Macrophage lipoprotein lipase modulates the development of atherosclerosis but not adiposity
    Article Snippet: Cross-sections of aortic roots were incubated with primary antibodies for mouse MOMA-2 (1:600; Accurate Chemical and Scientific) overnight at 4°C to investigate macrophage infiltration in the atherosclerotic lesion. .. After washing, sections were incubated with biotinylated anti-rat antibody for 1 h at room temperature, and then with avidin-biotin peroxidase complex (Vector Labs) for 30 min. Lastly, sections were developed with 3,3′-diaminobenzidine tetrahydrochloride (DAB) (Sigma), and counterstained with hematoxylin. .. The adipose tissue of mice fed a WTD for 12 weeks was fixed with 10% neutral-buffered formalin, embedded in paraffin, and sections were stained with hematoxylin and eosin.

    Article Title: Melatonin MT1 and MT2 Receptors in the Ram Reproductive Tract
    Article Snippet: The sections were then washed in PBS pH 7.4 and incubated in normal horse serum as a blocking reagent for 40 min, followed by overnight incubation at 4 °C with the specific primary antibody (rabbit polyclonal antibody against the MT1 receptor (RRID:AB_1241048, GeneTex, Irvine, CA, USA) and rabbit polyclonal antibody against the MT2 receptor (RRID:AB_1619198, Acris Antibodies GmbH, Herford, Germany), both at a dilution of 1:50 in PBS. .. Subsequently, the slides were incubated with biotinylated anti-rabbit antiserum (Vector Laboratories) for 30 min. An avidin-biotin-peroxidase complex (Vector Laboratories) was then applied for 40 min. .. The binding sites of the primary antibodies were visualized by diaminobenzidine (DAB) and peroxidase solution (0.12 g DAB in 240 mL of PBS pH 7.4 containing 3% H2 O2 ) for 7 min.

    Plasmid Preparation:

    Article Title: Anti-α4β1 Integrin Antibodies Attenuated Brain Inflammatory Changes in a Mouse Model of Alzheimer’s Disease
    Article Snippet: All mouse cytokine ELISA kits were obtained from R & D Systems (Minneapolis, MN). .. Elite Vectastain ABC avidin and biotin kits, biotinylated anti-rabbit, anti-mouse, and anti-rat antibodies and the Vector VIP kits were from Vector Laboratories Inc. (Burlingame, CA). .. Anti-CD68 was obtained from Serotec (Raleigh, NC).

    Staining:

    Article Title: Immunohistochemical analysis of macroautophagy
    Article Snippet: Subsequently, the primary antibody was applied overnight in a damp box, followed by 2 × 5 min immersion in staining buffer. .. Next, the slides were incubated for 30 min with biotinylated anti-rabbit or anti-mouse immunoglobulins (Vectastain ABC Kit, Vector Laboratories), 1:200 diluted in staining buffer. .. In case the primary antibody was already biotinylated, blocking with normal serum and incubation with biotinylated secondary antibodies were omitted.

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  • 99
    Vector Laboratories biotinylated secondary goat anti mouse antibody
    Assessments based on enzyme-linked immunosorbent assay of bound Aβ 1-42 -biotin does not distinguish between elderly high-pathology controls and elderly subjects with mild ADD. ( A ) There was no difference between groups in the levels of overall Aβ 1-42 -biotin recovered from dissociated tissue, as measured using an indirect ELISA which only detects <t>biotinylated</t> Aβ (not significant [n.s.] by Mann–Whitney U test). Data expressed as picograms Aβ per nanogram of total measurable protein. ( B ) The ratio of the amount of Aβ 1-42 -biotin to the amount of Aβ 1-x as measured by sandwich ELISA was not different between groups (not significant [n.s.] by Mann–Whitney U test). ( C ) The ratio of Aβ Aβ 1-42 -biotin as measured by sandwich ELISA to the percent gray matter plaque coverage did not differ between groups (not significant [n.s.] by Mann–Whitney U test). (D) There was no difference between groups in the levels of overall Aβ 1-x recovered from dissociated tissue, as measured using an sandwich ELISA (not significant [n.s.] by Mann–Whitney U test).
    Biotinylated Secondary Goat Anti Mouse Antibody, supplied by Vector Laboratories, 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/biotinylated secondary goat anti mouse antibody/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated secondary goat anti mouse antibody - by Bioz Stars, 2021-04
    99/100 stars
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    99
    Vector Laboratories biotinylated secondary antibody
    Schematic of the different binding steps on LMA. F9 lysates or purified p53 protein, purified primary p53 antibody, <t>biotinylated</t> secondary antibody, and Cy3-labeled streptavidin were printed on spots i, ii, iii and iv on the LMA, respectively. A diagrammatic representation of the different binding steps performed for signal detection on LMA is shown.
    Biotinylated Secondary Antibody, supplied by Vector Laboratories, 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/biotinylated secondary antibody/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated secondary antibody - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier

    99
    Vector Laboratories biotinylated goat anti rabbit antibody
    Representative illustration of IL-1RII immunostaining in the human endometrium. Sections of endometrial tissue were incubated with mouse monoclonal anti-IL-1RII antibody ( A , proliferative day 13; B , secretory day 24; original magnification, ×68) or with an equivalent concentration of normal mouse IgGs ( C and D , respectively; original magnification, ×68). Sections were then incubated successively with <t>biotinylated</t> goat anti-mouse polyclonal antibody and avidin-biotinylated horseradish peroxidase complex. The immunoreaction was revealed with diaminobenzidine (brown staining) and hematoxylin was used for counterstaining (blue staining). Note the brown fine positive staining in stromal and epithelial cells (cellular staining) ( E–H ; original magnification, ×268), and the brown deposit ( arrow ) that is primarily located at the apical side of glandular ( E , secretory phase day 24) and surface ( F , secretory phase day 16) epithelium, or more spread within the glands lumen ( G , secretory phase day 16). Positive immunostaining is also detected in isolated stromal cells (c) ( G , secretory phase day 16) and microvessels (v) ( H , secretory phase day 24) found in the stroma in the secretory phase of the menstrual cycle. s = stroma, g = gland.
    Biotinylated Goat Anti Rabbit Antibody, supplied by Vector Laboratories, 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/biotinylated goat anti rabbit antibody/product/Vector Laboratories
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated goat anti rabbit antibody - by Bioz Stars, 2021-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    Assessments based on enzyme-linked immunosorbent assay of bound Aβ 1-42 -biotin does not distinguish between elderly high-pathology controls and elderly subjects with mild ADD. ( A ) There was no difference between groups in the levels of overall Aβ 1-42 -biotin recovered from dissociated tissue, as measured using an indirect ELISA which only detects biotinylated Aβ (not significant [n.s.] by Mann–Whitney U test). Data expressed as picograms Aβ per nanogram of total measurable protein. ( B ) The ratio of the amount of Aβ 1-42 -biotin to the amount of Aβ 1-x as measured by sandwich ELISA was not different between groups (not significant [n.s.] by Mann–Whitney U test). ( C ) The ratio of Aβ Aβ 1-42 -biotin as measured by sandwich ELISA to the percent gray matter plaque coverage did not differ between groups (not significant [n.s.] by Mann–Whitney U test). (D) There was no difference between groups in the levels of overall Aβ 1-x recovered from dissociated tissue, as measured using an sandwich ELISA (not significant [n.s.] by Mann–Whitney U test).

    Journal: PLoS ONE

    Article Title: Soluble amyloid-beta buffering by plaques in Alzheimer disease dementia versus high-pathology controls

    doi: 10.1371/journal.pone.0200251

    Figure Lengend Snippet: Assessments based on enzyme-linked immunosorbent assay of bound Aβ 1-42 -biotin does not distinguish between elderly high-pathology controls and elderly subjects with mild ADD. ( A ) There was no difference between groups in the levels of overall Aβ 1-42 -biotin recovered from dissociated tissue, as measured using an indirect ELISA which only detects biotinylated Aβ (not significant [n.s.] by Mann–Whitney U test). Data expressed as picograms Aβ per nanogram of total measurable protein. ( B ) The ratio of the amount of Aβ 1-42 -biotin to the amount of Aβ 1-x as measured by sandwich ELISA was not different between groups (not significant [n.s.] by Mann–Whitney U test). ( C ) The ratio of Aβ Aβ 1-42 -biotin as measured by sandwich ELISA to the percent gray matter plaque coverage did not differ between groups (not significant [n.s.] by Mann–Whitney U test). (D) There was no difference between groups in the levels of overall Aβ 1-x recovered from dissociated tissue, as measured using an sandwich ELISA (not significant [n.s.] by Mann–Whitney U test).

    Article Snippet: The following day, sections were washed 3x in TBS for 5 minutes each and incubated with a biotinylated secondary goat anti-mouse antibody at a 1:1000 dilution in TBS-X for 1 hour at room temperature (Vector Laboratories).

    Techniques: Enzyme-linked Immunosorbent Assay, Indirect ELISA, MANN-WHITNEY, Sandwich ELISA

    Immunohistochemical staining for HAP1 in the human gastrointestinal tract. HAP1 was expressed in the cytoplasm. a Esophagus: There are few glands in the mucosa of esophagus. HAP1-positive cells were concentrated in these glands. b Stomach: Immunopositive cells were densely distributed in the gastric glands. c Small intestine showed scattered distribution of HAP1-immunoreactive cells in the villi and the intestine glands. d Colon showed weak and diffuse HAP1 staining in the colon glands. Amplification: left panel (× 5), middle panel (× 20) and right panel (× 40)

    Journal: Digestive diseases and sciences

    Article Title: Expression and Localization of Huntingtin‑Associated Protein 1 (HAP1) in the Human Digestive System

    doi: 10.1007/s10620-018-5425-5

    Figure Lengend Snippet: Immunohistochemical staining for HAP1 in the human gastrointestinal tract. HAP1 was expressed in the cytoplasm. a Esophagus: There are few glands in the mucosa of esophagus. HAP1-positive cells were concentrated in these glands. b Stomach: Immunopositive cells were densely distributed in the gastric glands. c Small intestine showed scattered distribution of HAP1-immunoreactive cells in the villi and the intestine glands. d Colon showed weak and diffuse HAP1 staining in the colon glands. Amplification: left panel (× 5), middle panel (× 20) and right panel (× 40)

    Article Snippet: Sections were then incubated with a purified rabbit antibody to human HAP1 (1:1000, EM39) at 4 °C for 40–45 h, followed by incubation of biotinylated goat anti-rabbit IgG (1:200, Vector Laboratories, USA) at room temperature for 1 h and avidin–biotin complex (Vectastain Elite ABC Reagent, USA) at room temperature for 1 h. Primary and secondary antibodies were diluted with PBS containing 0.2% Triton X-100 and 2% NGS with 3% BSA.

    Techniques: Immunohistochemistry, Staining, Amplification

    Western blots of huntingtin-associated protein 1 (HAP1) in different parts of the human gastrointestinal tract. a The blot was cut to stripes that were immunoprobed with antibodies to HAP1 and β-actin. Gastrointestinal tract protein extract (60 μg/lane) showed prominent immunoreactive bands at 75 kD for HAP1 and 42 kD for β-actin. Note that the expression of HAP1 was the highest in stomach and lowest in small intestine. b Densitometric analysis of the ratio of HAP1 to β-actin in each lane on the Western blots. Each bar represents mean ± SD ( n = 3)

    Journal: Digestive diseases and sciences

    Article Title: Expression and Localization of Huntingtin‑Associated Protein 1 (HAP1) in the Human Digestive System

    doi: 10.1007/s10620-018-5425-5

    Figure Lengend Snippet: Western blots of huntingtin-associated protein 1 (HAP1) in different parts of the human gastrointestinal tract. a The blot was cut to stripes that were immunoprobed with antibodies to HAP1 and β-actin. Gastrointestinal tract protein extract (60 μg/lane) showed prominent immunoreactive bands at 75 kD for HAP1 and 42 kD for β-actin. Note that the expression of HAP1 was the highest in stomach and lowest in small intestine. b Densitometric analysis of the ratio of HAP1 to β-actin in each lane on the Western blots. Each bar represents mean ± SD ( n = 3)

    Article Snippet: Sections were then incubated with a purified rabbit antibody to human HAP1 (1:1000, EM39) at 4 °C for 40–45 h, followed by incubation of biotinylated goat anti-rabbit IgG (1:200, Vector Laboratories, USA) at room temperature for 1 h and avidin–biotin complex (Vectastain Elite ABC Reagent, USA) at room temperature for 1 h. Primary and secondary antibodies were diluted with PBS containing 0.2% Triton X-100 and 2% NGS with 3% BSA.

    Techniques: Western Blot, Expressing

    Expression of HAP1 in normal gastric mucosa and gastric polyps. a Endoscopic view of a gastric polyp. b Protein levels of normal gastric mucosa and gastric polyps were analyzed by Western blotting. β-actin served as a loading control. c Densitometric analysis of the expression of HAP1 showed no difference between normal gastric mucosa and stomach polyps. Each bar represents mean ± SD of values ( n = 3, P > 0.05)

    Journal: Digestive diseases and sciences

    Article Title: Expression and Localization of Huntingtin‑Associated Protein 1 (HAP1) in the Human Digestive System

    doi: 10.1007/s10620-018-5425-5

    Figure Lengend Snippet: Expression of HAP1 in normal gastric mucosa and gastric polyps. a Endoscopic view of a gastric polyp. b Protein levels of normal gastric mucosa and gastric polyps were analyzed by Western blotting. β-actin served as a loading control. c Densitometric analysis of the expression of HAP1 showed no difference between normal gastric mucosa and stomach polyps. Each bar represents mean ± SD of values ( n = 3, P > 0.05)

    Article Snippet: Sections were then incubated with a purified rabbit antibody to human HAP1 (1:1000, EM39) at 4 °C for 40–45 h, followed by incubation of biotinylated goat anti-rabbit IgG (1:200, Vector Laboratories, USA) at room temperature for 1 h and avidin–biotin complex (Vectastain Elite ABC Reagent, USA) at room temperature for 1 h. Primary and secondary antibodies were diluted with PBS containing 0.2% Triton X-100 and 2% NGS with 3% BSA.

    Techniques: Expressing, Western Blot

    Comparison of HAP1 expression in pancreatitis, normal pancreas, and pancreatic cancer tissues. a Immunohistochemical staining of HAP1-positive cells in pancreatitis, normal pancreas, and pancreatic cancer tissue. Scale bar: 10 μm. b The head of the pancreas had been resected with Whipple procedure, and the pathology showed adenocarcinoma. c The expression of HAP1 in pancreatitis, normal pancreas, and pancreatic cancer tissue was detected by Western blotting. β-actin was a loading control. d Densitometric analysis showed the expression of HAP1 in pancreatitis, normal pancreas, and pancreatic cancer tissue ( n = 3, * P

    Journal: Digestive diseases and sciences

    Article Title: Expression and Localization of Huntingtin‑Associated Protein 1 (HAP1) in the Human Digestive System

    doi: 10.1007/s10620-018-5425-5

    Figure Lengend Snippet: Comparison of HAP1 expression in pancreatitis, normal pancreas, and pancreatic cancer tissues. a Immunohistochemical staining of HAP1-positive cells in pancreatitis, normal pancreas, and pancreatic cancer tissue. Scale bar: 10 μm. b The head of the pancreas had been resected with Whipple procedure, and the pathology showed adenocarcinoma. c The expression of HAP1 in pancreatitis, normal pancreas, and pancreatic cancer tissue was detected by Western blotting. β-actin was a loading control. d Densitometric analysis showed the expression of HAP1 in pancreatitis, normal pancreas, and pancreatic cancer tissue ( n = 3, * P

    Article Snippet: Sections were then incubated with a purified rabbit antibody to human HAP1 (1:1000, EM39) at 4 °C for 40–45 h, followed by incubation of biotinylated goat anti-rabbit IgG (1:200, Vector Laboratories, USA) at room temperature for 1 h and avidin–biotin complex (Vectastain Elite ABC Reagent, USA) at room temperature for 1 h. Primary and secondary antibodies were diluted with PBS containing 0.2% Triton X-100 and 2% NGS with 3% BSA.

    Techniques: Expressing, Immunohistochemistry, Staining, Western Blot

    Schematic of the different binding steps on LMA. F9 lysates or purified p53 protein, purified primary p53 antibody, biotinylated secondary antibody, and Cy3-labeled streptavidin were printed on spots i, ii, iii and iv on the LMA, respectively. A diagrammatic representation of the different binding steps performed for signal detection on LMA is shown.

    Journal: Proteome Science

    Article Title: Application of protein lysate microarrays to molecular marker verification and quantification

    doi: 10.1186/1477-5956-3-9

    Figure Lengend Snippet: Schematic of the different binding steps on LMA. F9 lysates or purified p53 protein, purified primary p53 antibody, biotinylated secondary antibody, and Cy3-labeled streptavidin were printed on spots i, ii, iii and iv on the LMA, respectively. A diagrammatic representation of the different binding steps performed for signal detection on LMA is shown.

    Article Snippet: Next, 50 μl buffer (50 mM Tris pH 7.5, 1.5 M NaCl, 0.1% Tween-20) containing biotinylated secondary antibody (Vector Laboratories, Burlingame, CA) at a concentration of 5 ng/μl was added to the arrays and a cover slip was placed on the arrays.

    Techniques: Binding Assay, Purification, Labeling

    Linearity of different binding steps on LMA. LMA containing serially diluted F9 protein lysates (0.055, 0.11, 0.23, 0.46, 0.92, 1.84, 3.68 and 7.36 ng), purified p53 protein (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), purified primary antibody (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), biotinylated secondary antibody (11.5, 23, 46, 92, 184, 368, 736 and 1472 pg) and Cy3-labeled streptavidin (1.95, 3.9, 7.8, 15.6, 31.2, 62.4, 124.8 and 249.6 ng) were printed. Arrays were probed with p53 antibody followed by binding and labeling steps as described in the Methods. Log transformed (base 2) spotted lysate amount was plotted against log transformed (base 2) signal intensity.

    Journal: Proteome Science

    Article Title: Application of protein lysate microarrays to molecular marker verification and quantification

    doi: 10.1186/1477-5956-3-9

    Figure Lengend Snippet: Linearity of different binding steps on LMA. LMA containing serially diluted F9 protein lysates (0.055, 0.11, 0.23, 0.46, 0.92, 1.84, 3.68 and 7.36 ng), purified p53 protein (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), purified primary antibody (1.5, 3, 6, 12, 24, 48, 96 and 192 pg), biotinylated secondary antibody (11.5, 23, 46, 92, 184, 368, 736 and 1472 pg) and Cy3-labeled streptavidin (1.95, 3.9, 7.8, 15.6, 31.2, 62.4, 124.8 and 249.6 ng) were printed. Arrays were probed with p53 antibody followed by binding and labeling steps as described in the Methods. Log transformed (base 2) spotted lysate amount was plotted against log transformed (base 2) signal intensity.

    Article Snippet: Next, 50 μl buffer (50 mM Tris pH 7.5, 1.5 M NaCl, 0.1% Tween-20) containing biotinylated secondary antibody (Vector Laboratories, Burlingame, CA) at a concentration of 5 ng/μl was added to the arrays and a cover slip was placed on the arrays.

    Techniques: Binding Assay, Purification, Labeling, Transformation Assay

    Representative illustration of IL-1RII immunostaining in the human endometrium. Sections of endometrial tissue were incubated with mouse monoclonal anti-IL-1RII antibody ( A , proliferative day 13; B , secretory day 24; original magnification, ×68) or with an equivalent concentration of normal mouse IgGs ( C and D , respectively; original magnification, ×68). Sections were then incubated successively with biotinylated goat anti-mouse polyclonal antibody and avidin-biotinylated horseradish peroxidase complex. The immunoreaction was revealed with diaminobenzidine (brown staining) and hematoxylin was used for counterstaining (blue staining). Note the brown fine positive staining in stromal and epithelial cells (cellular staining) ( E–H ; original magnification, ×268), and the brown deposit ( arrow ) that is primarily located at the apical side of glandular ( E , secretory phase day 24) and surface ( F , secretory phase day 16) epithelium, or more spread within the glands lumen ( G , secretory phase day 16). Positive immunostaining is also detected in isolated stromal cells (c) ( G , secretory phase day 16) and microvessels (v) ( H , secretory phase day 24) found in the stroma in the secretory phase of the menstrual cycle. s = stroma, g = gland.

    Journal: The American Journal of Pathology

    Article Title: Decreased Expression of the Decoy Interleukin-1 Receptor Type II in Human Endometriosis

    doi:

    Figure Lengend Snippet: Representative illustration of IL-1RII immunostaining in the human endometrium. Sections of endometrial tissue were incubated with mouse monoclonal anti-IL-1RII antibody ( A , proliferative day 13; B , secretory day 24; original magnification, ×68) or with an equivalent concentration of normal mouse IgGs ( C and D , respectively; original magnification, ×68). Sections were then incubated successively with biotinylated goat anti-mouse polyclonal antibody and avidin-biotinylated horseradish peroxidase complex. The immunoreaction was revealed with diaminobenzidine (brown staining) and hematoxylin was used for counterstaining (blue staining). Note the brown fine positive staining in stromal and epithelial cells (cellular staining) ( E–H ; original magnification, ×268), and the brown deposit ( arrow ) that is primarily located at the apical side of glandular ( E , secretory phase day 24) and surface ( F , secretory phase day 16) epithelium, or more spread within the glands lumen ( G , secretory phase day 16). Positive immunostaining is also detected in isolated stromal cells (c) ( G , secretory phase day 16) and microvessels (v) ( H , secretory phase day 24) found in the stroma in the secretory phase of the menstrual cycle. s = stroma, g = gland.

    Article Snippet: After a PBS rinse, the sections were incubated for 60 minutes with a rabbit polyclonal anti-IL-1β antibody diluted 8:1,000 in PBS-1% bovine serum albumin (R and D Systems), washed in PBS, incubated for 60 minutes with a biotinylated goat anti-rabbit antibody (Vector Laboratories) diluted 1:100 in PBS-1% bovine serum albumin, washed again in PBS, and finally incubated simultaneously for 60 minutes in the dark with fluorescein isothiocyanate-conjugated streptavidin and a rhodamine-conjugated goat anti-mouse antibody (Sigma), which were used at a final dilution of 1:100 and 1:10 in PBS-1% bovine serum albumin, respectively.

    Techniques: Immunostaining, Incubation, Concentration Assay, Avidin-Biotin Assay, Staining, Isolation