goat anti rabbit igg fitc conjugated  (Millipore)


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    Structured Review

    Millipore goat anti rabbit igg fitc conjugated
    Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary <t>anti-IgG-HRP</t> antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary <t>anti-IgG-FITC</t> antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.
    Goat Anti Rabbit Igg Fitc Conjugated, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 652 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Effects of Alcohol on Histone Deacetylase 2 (HDAC2) and the Neuroprotective Role of Trichostatin A (TSA)"

    Article Title: Effects of Alcohol on Histone Deacetylase 2 (HDAC2) and the Neuroprotective Role of Trichostatin A (TSA)

    Journal: Alcoholism, clinical and experimental research

    doi: 10.1111/j.1530-0277.2011.01492.x

    Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary anti-IgG-FITC antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.
    Figure Legend Snippet: Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary anti-IgG-FITC antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.

    Techniques Used: Incubation, Western Blot, Expressing, Flow Cytometry, Cytometry, Staining

    2) Product Images from "Vitronectins produced by human cirrhotic liver and CCl4‐treated rats differ in their glycosylation pattern and tissue remodeling activity"

    Article Title: Vitronectins produced by human cirrhotic liver and CCl4‐treated rats differ in their glycosylation pattern and tissue remodeling activity

    Journal: FEBS Open Bio

    doi: 10.1002/2211-5463.12616

    Amounts of protein and VN in CC l 4 ‐treated rat plasma. CC l 4 and olive oil were administered to rats twice a week. After 6 weeks, rats were sacrificed and blood collected. (A) Average weight of rats. Body weights of rats administered olive oil as a control (closed circle) and CC l 4 (open square) were measured at the time of administration. n = 6–13. (B) Total protein. Diluted plasma (100–200 times) was measured by using tunein‐ TP . Control: green bar; CC l 4 : yellow bar. n = 4. (C) VN level in plasma. Diluted plasma (2000–8000 times) was dot‐blotted onto a PVDF membrane and immunostained for VN . The staining intensity was measured by a refractive densitometer at 450 nm and corrected using the immunoreactivity of each VN . Control: green bar; CC l 4 : yellow bar. n = 4. (D) SDS / PAGE and immunostaining of VN s. Purified VN s (3 μg) from control and CC l 4 ‐treated plasma were loaded on each lane of a 9.5% acrylamide gel, and SDS / PAGE was performed in the presence of 2‐mercaptoethanol. Loading gels were transferred to PVDF membranes and stained with CBB (left) or sheep anti‐ VN IgG and HRP ‐anti‐sheep IgGs (right) as described in Materials and methods . Data are presented as mean ± SD . * P
    Figure Legend Snippet: Amounts of protein and VN in CC l 4 ‐treated rat plasma. CC l 4 and olive oil were administered to rats twice a week. After 6 weeks, rats were sacrificed and blood collected. (A) Average weight of rats. Body weights of rats administered olive oil as a control (closed circle) and CC l 4 (open square) were measured at the time of administration. n = 6–13. (B) Total protein. Diluted plasma (100–200 times) was measured by using tunein‐ TP . Control: green bar; CC l 4 : yellow bar. n = 4. (C) VN level in plasma. Diluted plasma (2000–8000 times) was dot‐blotted onto a PVDF membrane and immunostained for VN . The staining intensity was measured by a refractive densitometer at 450 nm and corrected using the immunoreactivity of each VN . Control: green bar; CC l 4 : yellow bar. n = 4. (D) SDS / PAGE and immunostaining of VN s. Purified VN s (3 μg) from control and CC l 4 ‐treated plasma were loaded on each lane of a 9.5% acrylamide gel, and SDS / PAGE was performed in the presence of 2‐mercaptoethanol. Loading gels were transferred to PVDF membranes and stained with CBB (left) or sheep anti‐ VN IgG and HRP ‐anti‐sheep IgGs (right) as described in Materials and methods . Data are presented as mean ± SD . * P

    Techniques Used: Staining, SDS Page, Immunostaining, Purification, Acrylamide Gel Assay

    3) Product Images from "A Novel View on the Role of Intracellular Tails in Surface Delivery of the Potassium-Chloride Cotransporter KCC2"

    Article Title: A Novel View on the Role of Intracellular Tails in Surface Delivery of the Potassium-Chloride Cotransporter KCC2

    Journal: eNeuro

    doi: 10.1523/ENEURO.0055-17.2017

    Visualization of surface expressed and internalized KCC2-pH ext proteins using a live-cell immunolabeling protocol on cultured hippocampal neurons. A , Scheme of the multistep immunolabeling protocol applied to 13 DIV neurons. First Ab, primary antibody; second Cy3, Cy3-conjugated secondary antibody; second Alexa 647, Alexa Fluor 647–conjugated secondary antibody; PFA, paraformaldehyde. The scheme does not include final steps of fixed and permeabilized cells labeled with mouse anti-GFP and anti-mouse Alexa Fluor 488 antibody [total protein pool (F t )]. B , Representative images showing fluorescence emitted after staining with Cy3-conjugated secondary antibody [plasma membrane restricted pool (F m )]; images were pseudocolored using illustrated bicolor lookup table, first raw); Alexa Fluor 647–conjugated secondary antibody (internalized surface labeled molecules and portion of surface retained molecules, second raw); Alexa Fluor 488–conjugated secondary antibody (F t , third raw); internalized surface labeled signal obtained by arithmetic subtraction of first and second raw images (F i , fourth raw). Image columns illustrate fluorescent signals obtained at different z -planes or after arithmetic summation of nine planes as indicated. The neuronal shape (Alexa Fluor 488 fluorescence) is shown in light green in each image for reference. Insets illustrate indicated portions of images at higher zoom. Scale bars: 8 µm (main image), 1 µm (insets).
    Figure Legend Snippet: Visualization of surface expressed and internalized KCC2-pH ext proteins using a live-cell immunolabeling protocol on cultured hippocampal neurons. A , Scheme of the multistep immunolabeling protocol applied to 13 DIV neurons. First Ab, primary antibody; second Cy3, Cy3-conjugated secondary antibody; second Alexa 647, Alexa Fluor 647–conjugated secondary antibody; PFA, paraformaldehyde. The scheme does not include final steps of fixed and permeabilized cells labeled with mouse anti-GFP and anti-mouse Alexa Fluor 488 antibody [total protein pool (F t )]. B , Representative images showing fluorescence emitted after staining with Cy3-conjugated secondary antibody [plasma membrane restricted pool (F m )]; images were pseudocolored using illustrated bicolor lookup table, first raw); Alexa Fluor 647–conjugated secondary antibody (internalized surface labeled molecules and portion of surface retained molecules, second raw); Alexa Fluor 488–conjugated secondary antibody (F t , third raw); internalized surface labeled signal obtained by arithmetic subtraction of first and second raw images (F i , fourth raw). Image columns illustrate fluorescent signals obtained at different z -planes or after arithmetic summation of nine planes as indicated. The neuronal shape (Alexa Fluor 488 fluorescence) is shown in light green in each image for reference. Insets illustrate indicated portions of images at higher zoom. Scale bars: 8 µm (main image), 1 µm (insets).

    Techniques Used: Immunolabeling, Cell Culture, Labeling, Fluorescence, Staining

    4) Product Images from "Interaction of Gene-Cloned and Insect Cell-Expressed Aminopeptidase N of Spodoptera litura with Insecticidal Crystal Protein Cry1C"

    Article Title: Interaction of Gene-Cloned and Insect Cell-Expressed Aminopeptidase N of Spodoptera litura with Insecticidal Crystal Protein Cry1C

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.68.9.4583-4592.2002

    Immunolocalization of expressed S. litura APN. Surface expression of the APN was examined at 36 h postinfection by immunofluorescence staining with rabbit polyclonal antibodies raised against APN and FITC-conjugated goat anti-rabbit IgG. (A) Cells overlaid with preimmune serum. (B) Cells overlaid with anti-APN antibodies.
    Figure Legend Snippet: Immunolocalization of expressed S. litura APN. Surface expression of the APN was examined at 36 h postinfection by immunofluorescence staining with rabbit polyclonal antibodies raised against APN and FITC-conjugated goat anti-rabbit IgG. (A) Cells overlaid with preimmune serum. (B) Cells overlaid with anti-APN antibodies.

    Techniques Used: Expressing, Immunofluorescence, Staining

    Ligand blot analysis of insect cells expressing S. litura APN. Membrane proteins (2.5 μg) prepared from uninfected Sf21 cells (lane 1) and BV-SlApn-infected cells (lane 2 and 3) were solubilized in SDS sample buffer containing β-mercaptoethanol, separated by SDS-7.5% PAGE, and electrotransferred to nitrocellulose membrane. The nitrocellulose membrane was probed with Cry1C (1 μg/ml) (lanes 1 and 2) and Cry1Ac (1 μg/ml) (lane 3) δ-endotoxins. Binding of the toxin was detected by using antibodies against the toxin followed by alkaline phosphatase-conjugated goat anti-rabbit IgG. The positions of molecular mass markers (in kilodaltons) are indicated on the left.
    Figure Legend Snippet: Ligand blot analysis of insect cells expressing S. litura APN. Membrane proteins (2.5 μg) prepared from uninfected Sf21 cells (lane 1) and BV-SlApn-infected cells (lane 2 and 3) were solubilized in SDS sample buffer containing β-mercaptoethanol, separated by SDS-7.5% PAGE, and electrotransferred to nitrocellulose membrane. The nitrocellulose membrane was probed with Cry1C (1 μg/ml) (lanes 1 and 2) and Cry1Ac (1 μg/ml) (lane 3) δ-endotoxins. Binding of the toxin was detected by using antibodies against the toxin followed by alkaline phosphatase-conjugated goat anti-rabbit IgG. The positions of molecular mass markers (in kilodaltons) are indicated on the left.

    Techniques Used: Expressing, Infection, Polyacrylamide Gel Electrophoresis, Binding Assay

    Differential toxin binding to surface-expressed S. litura APN. Sf21 cells infected for 36 h were overlaid with Cry1Ac (A) or Cry1C (B) toxin (1 μg/ml). Binding of the toxin was detected by using antibodies against the toxin followed by FITC-conjugated goat anti-rabbit IgG.
    Figure Legend Snippet: Differential toxin binding to surface-expressed S. litura APN. Sf21 cells infected for 36 h were overlaid with Cry1Ac (A) or Cry1C (B) toxin (1 μg/ml). Binding of the toxin was detected by using antibodies against the toxin followed by FITC-conjugated goat anti-rabbit IgG.

    Techniques Used: Binding Assay, Infection

    5) Product Images from "A multivalent Kaposi sarcoma-associated herpesvirus-like particle vaccine capable of eliciting high titers of neutralizing antibodies in immunized rabbits"

    Article Title: A multivalent Kaposi sarcoma-associated herpesvirus-like particle vaccine capable of eliciting high titers of neutralizing antibodies in immunized rabbits

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2019.04.071

    KSHV-glycoprotein-specific IgG titers in immunized New Zealand white rabbits. (a) Coomassie stain (left) and immunoblot (right) of KSHV gpK8.1, gB, and gH/gL recombinant Fc-His tagged proteins. Fc-6xHis-tagged recombinant KSHV gpK8.1, gB, and gH/gL proteins were expressed by transiently transfecting HEK-293–6E cells. Culture media was harvested six days post-transfection by centrifugation and filtration through a 0.22 µM. Fc-6xHis-tagged KSHV proteins in the media were purified using protein A spin-columns, concentrated in PBS using Amicon Ultra 15 centrifugal filter units, and quantified using a nanodrop spectrophotometer. To confirm the specificity of the proteins, the concentrated proteins were separated on a 4–12% SDS-PAGE and detected by Coomassie blue stain (for molecular weight) or transferred to a polyvinylidene fluoride membrane for immunoblot analysis using monoclonal anti-gpK8.1 or anti-gH/gL or polyclonal goat anti-human Fc (gB) antibodies as indicated. (b) Immunization and bleeding schedules of wild-type New Zealand white rabbits. Eight- to 10-week-old rabbits (n = 6/treatment) were immunized subcutaneously at Days 0, 28 and 42 with 50 µg purified KSHV-LPs−/+HR2, UV-inactivated KSHV, or TNE buffer, all adsorbed to alum and MPL as adjuvants. Immunized rabbits were bled seven days pre-immunization (−7) and on Days 14, 35, 49, 70, and 90 (terminal bleed). (c) Serum KSHV glycoprotein-specific antibody responses. KSHV-glycoprotein IgG-specific antibody titers in diluted (1:300 and 1:900) sera from immunized rabbits were determined using ELISA with recombinant tagged gpK8.1, gB, and gH/gL proteins; results of quadruplicate replicates for each of the six animals per group are expressed as mean ± standard deviation (SD). Differences in antibody titers between all groups were analyzed using a Kruskal-Wallis test; differences between the −HR2 and+HR2 vaccine were assessed using a Mann-Whitney test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
    Figure Legend Snippet: KSHV-glycoprotein-specific IgG titers in immunized New Zealand white rabbits. (a) Coomassie stain (left) and immunoblot (right) of KSHV gpK8.1, gB, and gH/gL recombinant Fc-His tagged proteins. Fc-6xHis-tagged recombinant KSHV gpK8.1, gB, and gH/gL proteins were expressed by transiently transfecting HEK-293–6E cells. Culture media was harvested six days post-transfection by centrifugation and filtration through a 0.22 µM. Fc-6xHis-tagged KSHV proteins in the media were purified using protein A spin-columns, concentrated in PBS using Amicon Ultra 15 centrifugal filter units, and quantified using a nanodrop spectrophotometer. To confirm the specificity of the proteins, the concentrated proteins were separated on a 4–12% SDS-PAGE and detected by Coomassie blue stain (for molecular weight) or transferred to a polyvinylidene fluoride membrane for immunoblot analysis using monoclonal anti-gpK8.1 or anti-gH/gL or polyclonal goat anti-human Fc (gB) antibodies as indicated. (b) Immunization and bleeding schedules of wild-type New Zealand white rabbits. Eight- to 10-week-old rabbits (n = 6/treatment) were immunized subcutaneously at Days 0, 28 and 42 with 50 µg purified KSHV-LPs−/+HR2, UV-inactivated KSHV, or TNE buffer, all adsorbed to alum and MPL as adjuvants. Immunized rabbits were bled seven days pre-immunization (−7) and on Days 14, 35, 49, 70, and 90 (terminal bleed). (c) Serum KSHV glycoprotein-specific antibody responses. KSHV-glycoprotein IgG-specific antibody titers in diluted (1:300 and 1:900) sera from immunized rabbits were determined using ELISA with recombinant tagged gpK8.1, gB, and gH/gL proteins; results of quadruplicate replicates for each of the six animals per group are expressed as mean ± standard deviation (SD). Differences in antibody titers between all groups were analyzed using a Kruskal-Wallis test; differences between the −HR2 and+HR2 vaccine were assessed using a Mann-Whitney test. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Techniques Used: Staining, Recombinant, Transfection, Centrifugation, Filtration, Purification, Spectrophotometry, SDS Page, Molecular Weight, Enzyme-linked Immunosorbent Assay, Standard Deviation, MANN-WHITNEY

    6) Product Images from "Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG against Leptospira for Early Detection of Acute Leptospirosis"

    Article Title: Production and Characterization of a Polyclonal Antibody of Anti-rLipL21-IgG against Leptospira for Early Detection of Acute Leptospirosis

    Journal: BioMed Research International

    doi: 10.1155/2014/592858

    Purification of (anti-rLipL21) IgG antibody was purified from rabbit antiserum against rLipL21. Lane L: anti-rLipL21 serum; Lane FT: flow through of unbound proteins from column; Lane W: wash of nonspecific proteins from column; Lane 1: eluted fraction without protein; Lane 2: purified anti-rLipL21-IgG; Lane 3: concentrated anti-rLipL21-IgG. Molecular weight standards are indicated in kilodaltons.
    Figure Legend Snippet: Purification of (anti-rLipL21) IgG antibody was purified from rabbit antiserum against rLipL21. Lane L: anti-rLipL21 serum; Lane FT: flow through of unbound proteins from column; Lane W: wash of nonspecific proteins from column; Lane 1: eluted fraction without protein; Lane 2: purified anti-rLipL21-IgG; Lane 3: concentrated anti-rLipL21-IgG. Molecular weight standards are indicated in kilodaltons.

    Techniques Used: Purification, Flow Cytometry, Molecular Weight

    Immunoblot of panel of Leptospira spp. obtained by using rabbit anti-rLipL21-IgG antibody to detect single band in leptospiral LipL21 antigen in detergent phase. Lane 1: Canicola (strain Hond Utrecht IV); Lane 2: Hardjobovis (Sponselee); Lane 3: Autumnalis (strain Akiyami A); Lane 4: Icterohaemorrhagiae (strain RGA); Lane 5: Australis (strain Ballico); Lane 6: Pomona (strain Pomona); Lane 7: Grippotyphosa (strain Moskva V); Lane 8: L. kmetyi serovar Malaysia strain Bejo-iso 9 T ; Lane 9: Balum (strain Mus 127); Lane 10: Grippotyphosa (strain Moskva V); Lane 11: Hebdomadis (strain Hebdomadis); Lane 12: nonpathogenic species L. biflexa (strain Patoc 1). Molecular weight standards are indicated in kilodaltons.
    Figure Legend Snippet: Immunoblot of panel of Leptospira spp. obtained by using rabbit anti-rLipL21-IgG antibody to detect single band in leptospiral LipL21 antigen in detergent phase. Lane 1: Canicola (strain Hond Utrecht IV); Lane 2: Hardjobovis (Sponselee); Lane 3: Autumnalis (strain Akiyami A); Lane 4: Icterohaemorrhagiae (strain RGA); Lane 5: Australis (strain Ballico); Lane 6: Pomona (strain Pomona); Lane 7: Grippotyphosa (strain Moskva V); Lane 8: L. kmetyi serovar Malaysia strain Bejo-iso 9 T ; Lane 9: Balum (strain Mus 127); Lane 10: Grippotyphosa (strain Moskva V); Lane 11: Hebdomadis (strain Hebdomadis); Lane 12: nonpathogenic species L. biflexa (strain Patoc 1). Molecular weight standards are indicated in kilodaltons.

    Techniques Used: Molecular Weight

    7) Product Images from "A Well-Connected and Conserved Nucleoplasmic Helicase Is Required for Production of Box C/D and H/ACA snoRNAs and Localization of snoRNP Proteins"

    Article Title: A Well-Connected and Conserved Nucleoplasmic Helicase Is Required for Production of Box C/D and H/ACA snoRNAs and Localization of snoRNP Proteins

    Journal: Molecular and Cellular Biology

    doi: 10.1128/MCB.21.22.7731-7746.2001

    Kinetics of Rvb2 depletion in galactose-dependent test cells. Test cells dependent on a GAL :: rvb2 allele (strain YWD313) were grown in medium containing 2% galactose, washed, and shifted to medium containing 2% glucose. Cell samples were collected before and at various intervals after the medium shift. (A) Northern analysis of RVB2 mRNA accumulation. Total RNA was extracted from cells at the time points indicated and fractionated on an agarose-formaldehyde gel, and a blot was probed with radiolabeled DNA that recognizes RVB2 mRNA. (B) Western analysis of Rvb2p protein accumulation. Total protein was extracted at the time points indicated, fractionated by SDS-polyacrylamide gel electrophoresis, and electroblotted to a nitrocellulose membrane. Proteins were detected with rabbit polyclonal IgG (raised against recombinant Rvb2p) and a goat anti-rabbit–peroxidase conjugate. Left lane, total protein from a strain (YWD312) carrying HIS3 :: rvb2 and Rvb2-GFP expressed from a plasmid; the other lanes show total proteins from GAL :: rvb2 cells incubated in glucose for 0, 2, and 8 h, as indicated.
    Figure Legend Snippet: Kinetics of Rvb2 depletion in galactose-dependent test cells. Test cells dependent on a GAL :: rvb2 allele (strain YWD313) were grown in medium containing 2% galactose, washed, and shifted to medium containing 2% glucose. Cell samples were collected before and at various intervals after the medium shift. (A) Northern analysis of RVB2 mRNA accumulation. Total RNA was extracted from cells at the time points indicated and fractionated on an agarose-formaldehyde gel, and a blot was probed with radiolabeled DNA that recognizes RVB2 mRNA. (B) Western analysis of Rvb2p protein accumulation. Total protein was extracted at the time points indicated, fractionated by SDS-polyacrylamide gel electrophoresis, and electroblotted to a nitrocellulose membrane. Proteins were detected with rabbit polyclonal IgG (raised against recombinant Rvb2p) and a goat anti-rabbit–peroxidase conjugate. Left lane, total protein from a strain (YWD312) carrying HIS3 :: rvb2 and Rvb2-GFP expressed from a plasmid; the other lanes show total proteins from GAL :: rvb2 cells incubated in glucose for 0, 2, and 8 h, as indicated.

    Techniques Used: Northern Blot, Western Blot, Polyacrylamide Gel Electrophoresis, Recombinant, Plasmid Preparation, Incubation

    8) Product Images from "Effects of Alcohol on Histone Deacetylase 2 (HDAC2) and the Neuroprotective Role of Trichostatin A (TSA)"

    Article Title: Effects of Alcohol on Histone Deacetylase 2 (HDAC2) and the Neuroprotective Role of Trichostatin A (TSA)

    Journal: Alcoholism, clinical and experimental research

    doi: 10.1111/j.1530-0277.2011.01492.x

    Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary anti-IgG-FITC antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.
    Figure Legend Snippet: Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary anti-IgG-FITC antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.

    Techniques Used: Incubation, Western Blot, Expressing, Flow Cytometry, Cytometry, Staining

    9) Product Images from "Alcohol-induced serotonergic modulation: the role of histone deacetylases"

    Article Title: Alcohol-induced serotonergic modulation: the role of histone deacetylases

    Journal: Alcohol (Fayetteville, N.Y.)

    doi: 10.1016/j.alcohol.2012.03.005

    TSA significantly enhances ethanol effects on 5-HT3. After reaching 60% confluency, SK-N-MC and human primary neurons were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 and 72 hrs. Gene expression (48 hrs) was performed in both primary neurons and SK-N-MC (figure 4a) as described above. Data are expressed as mean ± SEM of 5-HT3 gene expression of six independent experiments. Protein levels (72 hrs) were measured in SK-N-MC by western blot and flow cytometry. For the western blot experiments (figure 4b), 20 μg of protein from SK-N-MC whole cell lysates were stained with primary anti-5-HT3 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot. Protein quantification is expressed as % control ± SEM of 5-HT3 protein of six independent experiments. * represents significance compared to control (*** p
    Figure Legend Snippet: TSA significantly enhances ethanol effects on 5-HT3. After reaching 60% confluency, SK-N-MC and human primary neurons were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 and 72 hrs. Gene expression (48 hrs) was performed in both primary neurons and SK-N-MC (figure 4a) as described above. Data are expressed as mean ± SEM of 5-HT3 gene expression of six independent experiments. Protein levels (72 hrs) were measured in SK-N-MC by western blot and flow cytometry. For the western blot experiments (figure 4b), 20 μg of protein from SK-N-MC whole cell lysates were stained with primary anti-5-HT3 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot. Protein quantification is expressed as % control ± SEM of 5-HT3 protein of six independent experiments. * represents significance compared to control (*** p

    Techniques Used: Incubation, Expressing, Western Blot, Flow Cytometry, Cytometry, Staining

    10) Product Images from "Selective Expression of CYP2A13 in Human Pancreatic ?-Islet Cells"

    Article Title: Selective Expression of CYP2A13 in Human Pancreatic ?-Islet Cells

    Journal: Drug Metabolism and Disposition

    doi: 10.1124/dmd.112.046359

    Immunofluorescent double staining images of CYP2A13, proinsulin C-peptide, and glucagon in adult human pancreatic islets (magnification, 400×). A-1 and B-1, rabbit anti-CYP2A13 antibody and FITC-conjugated goat anti-rabbit IgG were used as the
    Figure Legend Snippet: Immunofluorescent double staining images of CYP2A13, proinsulin C-peptide, and glucagon in adult human pancreatic islets (magnification, 400×). A-1 and B-1, rabbit anti-CYP2A13 antibody and FITC-conjugated goat anti-rabbit IgG were used as the

    Techniques Used: Double Staining

    11) Product Images from "Up-regulation of miR-21 Mediates Resistance to Trastuzumab Therapy for Breast Cancer *"

    Article Title: Up-regulation of miR-21 Mediates Resistance to Trastuzumab Therapy for Breast Cancer *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.216887

    Retrieving PTEN expression re-sensitizes breast cancer cells to trastuzumab. A , diagram of the p3xFLAG- PTEN -mut construct with three mutated nucleotides ( arrowheads ) at the miR-21 binding site within the 3′ UTR of PTEN. B , PTEN phosphatase activities ( upper ) and Western blotting for PTEN expression ( lower ) in the membrane precipitates with an anti-PTEN antibody or a control isotype IgG in the trastuzumab-resistant BT474 cells that were transfected with miR-21 ASO, Lin4 ASO ( left ), or with p3XFLAG- PTEN -MUT or empty p3XFLAG vector ( right ) and treated with trastuzumab for the indicated periods of time. C , Western blotting for the expression of the total protein and the phosphorylated protein of AKT at Ser-473 in the trastuzumab-resistant and -sensitive BT474 cells mock transfected or transfected with miR-21 ASO, Lin4 ASO, p3XFLAG- PTEN -mut, or p3XFLAG vector alone. D , [ 3 H]thymidine incorporation assays for the trastuzumab-resistant BT474 cells that were mock transfected (■) or transfected with p3XFLAG- PTEN -mut (●) or p3XFLAG vector alone (◊) and treated with increasing concentrations of trastuzumab; **, p
    Figure Legend Snippet: Retrieving PTEN expression re-sensitizes breast cancer cells to trastuzumab. A , diagram of the p3xFLAG- PTEN -mut construct with three mutated nucleotides ( arrowheads ) at the miR-21 binding site within the 3′ UTR of PTEN. B , PTEN phosphatase activities ( upper ) and Western blotting for PTEN expression ( lower ) in the membrane precipitates with an anti-PTEN antibody or a control isotype IgG in the trastuzumab-resistant BT474 cells that were transfected with miR-21 ASO, Lin4 ASO ( left ), or with p3XFLAG- PTEN -MUT or empty p3XFLAG vector ( right ) and treated with trastuzumab for the indicated periods of time. C , Western blotting for the expression of the total protein and the phosphorylated protein of AKT at Ser-473 in the trastuzumab-resistant and -sensitive BT474 cells mock transfected or transfected with miR-21 ASO, Lin4 ASO, p3XFLAG- PTEN -mut, or p3XFLAG vector alone. D , [ 3 H]thymidine incorporation assays for the trastuzumab-resistant BT474 cells that were mock transfected (■) or transfected with p3XFLAG- PTEN -mut (●) or p3XFLAG vector alone (◊) and treated with increasing concentrations of trastuzumab; **, p

    Techniques Used: Expressing, Construct, Binding Assay, Western Blot, Transfection, Allele-specific Oligonucleotide, Plasmid Preparation

    12) Product Images from "Role of Kinase Suppressor of Ras-1 in Neuronal Survival Signaling by Extracellular Signal-Regulated Kinase 1/2"

    Article Title: Role of Kinase Suppressor of Ras-1 in Neuronal Survival Signaling by Extracellular Signal-Regulated Kinase 1/2

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.3473-07.2007

    KSR1 associates with active ERK1/2 in BDNF-stimulated rat cortical neurons. A , KSR1 expression in the adult rat brain. KSR1 immunoreactivity in coronal rat brain sections was visualized by immunofluorescence with a rabbit anti-KSR1 antibody ( b′ , c′ , e′ , f′ ); IgG from nonimmunized rabbits was used to control the specificity of staining ( a′ , d′ ). Low-magnification views of the neocortex ( a′ , b′ ) and the hippocampus ( d′ , e′ ) are shown. In c′ and f′ , higher-magnification views of the neocortical layers 4/5 and the CA3 subfield of the hippocampus are presented as indicated. Single confocal sections are shown; a similar pattern of KSR1 staining was observed in sections from four different animals. B , Coimmunoprecipitation of KSR1 and active ERK1/2 from BDNF-stimulated neurons. Cortical neurons were treated as indicated. Proteins immunoprecipitated with a KSR1 antibody were analyzed by Western blotting for KSR1, active ERK1/2 (phosphorylated at Thr 183 and Tyr 185 , pERK1/2), total ERK1/2 (ERK1/2) or MKK1/2. BDNF induced the association between KSR1 and pERK1/2. Note the presence of total ERK1/2 associated with KSR1 in unstimulated neurons. No pERK1/2 or ERK1/2 was found if the immunoprecipitation (IP) was performed using nonimmune IgG (data not shown). The interactions between KSR1 and MKK1/2 were unaffected by BDNF. Similar levels of KSR1 indicated consistent immunoprecipitation. The graph presents quantitative analysis of pERK1/2-KSR1 association from three independent experiments. Error bars are SEM. p
    Figure Legend Snippet: KSR1 associates with active ERK1/2 in BDNF-stimulated rat cortical neurons. A , KSR1 expression in the adult rat brain. KSR1 immunoreactivity in coronal rat brain sections was visualized by immunofluorescence with a rabbit anti-KSR1 antibody ( b′ , c′ , e′ , f′ ); IgG from nonimmunized rabbits was used to control the specificity of staining ( a′ , d′ ). Low-magnification views of the neocortex ( a′ , b′ ) and the hippocampus ( d′ , e′ ) are shown. In c′ and f′ , higher-magnification views of the neocortical layers 4/5 and the CA3 subfield of the hippocampus are presented as indicated. Single confocal sections are shown; a similar pattern of KSR1 staining was observed in sections from four different animals. B , Coimmunoprecipitation of KSR1 and active ERK1/2 from BDNF-stimulated neurons. Cortical neurons were treated as indicated. Proteins immunoprecipitated with a KSR1 antibody were analyzed by Western blotting for KSR1, active ERK1/2 (phosphorylated at Thr 183 and Tyr 185 , pERK1/2), total ERK1/2 (ERK1/2) or MKK1/2. BDNF induced the association between KSR1 and pERK1/2. Note the presence of total ERK1/2 associated with KSR1 in unstimulated neurons. No pERK1/2 or ERK1/2 was found if the immunoprecipitation (IP) was performed using nonimmune IgG (data not shown). The interactions between KSR1 and MKK1/2 were unaffected by BDNF. Similar levels of KSR1 indicated consistent immunoprecipitation. The graph presents quantitative analysis of pERK1/2-KSR1 association from three independent experiments. Error bars are SEM. p

    Techniques Used: Expressing, Immunofluorescence, Staining, Immunoprecipitation, Western Blot

    13) Product Images from "Elevated expression of syntenin in breast cancer is correlated with lymph node metastasis and poor patient survival"

    Article Title: Elevated expression of syntenin in breast cancer is correlated with lymph node metastasis and poor patient survival

    Journal: Breast Cancer Research : BCR

    doi: 10.1186/bcr3442

    Activation of integrin β1 and ERK1/2 is essential for syntenin-induced migration and invasion . (A) Silencing of syntenin in MDA-MB-231HM cells inhibited active integrin β1 expression and phosphorylation of ERK1/2, but had no effects on JNK and p38. (B) Overexpression of syntenin increased active integrin β1 expression and ERK1/2 phosphorylation in 231-SYN cells. The activation of ERK1/2 was analyzed with Western blot by using phosphor-specific antibodies. (C) Integrin β1 functional blocking antibody blocked both active integrin β1 expression and ERK1/2 phosphorylation. Cells were treated with integrin β1 or nonspecific IgG for 1 hour before protein extraction. (D) ERK1/2 inhibitor U0126 blocked activation of ERK1/2. Cells were pretreated with dimethylsulfoxide (DMSO) or 20 µ M U0126 (U0126) for 2 hours before protein extraction. (E, F) U0126 effectively reduced the migration and invasion of breast cancer cells. GAPDH was used as loading control. Data are expressed as means of triplicate samples from three independent experiments; bars, SD. ** P
    Figure Legend Snippet: Activation of integrin β1 and ERK1/2 is essential for syntenin-induced migration and invasion . (A) Silencing of syntenin in MDA-MB-231HM cells inhibited active integrin β1 expression and phosphorylation of ERK1/2, but had no effects on JNK and p38. (B) Overexpression of syntenin increased active integrin β1 expression and ERK1/2 phosphorylation in 231-SYN cells. The activation of ERK1/2 was analyzed with Western blot by using phosphor-specific antibodies. (C) Integrin β1 functional blocking antibody blocked both active integrin β1 expression and ERK1/2 phosphorylation. Cells were treated with integrin β1 or nonspecific IgG for 1 hour before protein extraction. (D) ERK1/2 inhibitor U0126 blocked activation of ERK1/2. Cells were pretreated with dimethylsulfoxide (DMSO) or 20 µ M U0126 (U0126) for 2 hours before protein extraction. (E, F) U0126 effectively reduced the migration and invasion of breast cancer cells. GAPDH was used as loading control. Data are expressed as means of triplicate samples from three independent experiments; bars, SD. ** P

    Techniques Used: Activation Assay, Migration, Multiple Displacement Amplification, Expressing, Over Expression, Western Blot, Functional Assay, Blocking Assay, Protein Extraction

    Related Articles

    Centrifugation:

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    Filtration:

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    Blocking Assay:

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    Enzyme-linked Immunosorbent Assay:

    Article Title: Production of Native Bispecific Antibodies in Rabbits
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    Sandwich ELISA:

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    Incubation:

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    Article Title: Peculiar Expression of CD3-Epsilon in Kidney of Ginbuna Crucian Carp
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    Article Title: Production of Native Bispecific Antibodies in Rabbits
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    Article Title: Evaluation of YadC protein delivered by live attenuated Salmonella as a vaccine against plague
    Article Snippet: The membranes were blocked with 3% skim milk in PBS and incubated with rabbit polyclonal anti-YadC antibodies for 1 h at room temperature. .. Then, horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G (IgG) (Sigma) was added in 1X PBS/3% skim milk.

    Article Title: Accelerated immunoassays based on magnetic particle dynamics in a rotating capillary tube with stationary magnetic field
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    Infection:

    Article Title: LcrV of Yersinia pestis Enters Infected Eukaryotic Cells by a Virulence Plasmid-Independent Mechanism
    Article Snippet: Proteins from fractionated, infected HeLa cultures were resolved in 12% (wt/vol) polyacrylamide gels by SDS-PAGE ( ). .. Proteins were visualized by treatment of immunoblots with alkaline phosphatase- or horseradish peroxidase-conjugated goat anti-rabbit IgG (Sigma), followed by development with nitroblue tetrazolium–5-bromo-4-chloro-3-indolylphosphate (NBT-BCIP; GIBCO-BRL) or enhanced chemiluminescence (ECL) substrate (Pierce), respectively.

    Bradford Assay:

    Article Title: Adenylate Cyclase Toxin from Bordetella pertussis Synergizes with Lipopolysaccharide To Promote Innate Interleukin-10 Production and Enhances the Induction of Th2 and Regulatory T Cells
    Article Snippet: LPS was measured by a colorimetric Limulus amoebocyte lysate assay (QCL-1000; Biowhittaker, Walkersville, Md.), and protein concentrations were determined by Bradford assay (Bio-Rad). .. The bands were visualized by incubation with secondary anti-rabbit immunoglobulin G (IgG) horseradish peroxidase-conjugated antibodies (Sigma) and chemiluminescent supersignal detection system (Pierce).

    Western Blot:

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    Article Title: Peculiar Expression of CD3-Epsilon in Kidney of Ginbuna Crucian Carp
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    Article Title: Display of Bacterial Lipase on the Escherichia coli Cell Surface by Using FadL as an Anchoring Motif and Use of the Enzyme in Enantioselective Biocatalysis
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    Article Title: Evaluation of YadC protein delivered by live attenuated Salmonella as a vaccine against plague
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    Article Title: An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles
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    Article Title: LcrV of Yersinia pestis Enters Infected Eukaryotic Cells by a Virulence Plasmid-Independent Mechanism
    Article Snippet: .. Proteins were visualized by treatment of immunoblots with alkaline phosphatase- or horseradish peroxidase-conjugated goat anti-rabbit IgG (Sigma), followed by development with nitroblue tetrazolium–5-bromo-4-chloro-3-indolylphosphate (NBT-BCIP; GIBCO-BRL) or enhanced chemiluminescence (ECL) substrate (Pierce), respectively. .. A tissue culture infection model has been used to characterize the localization and function of certain Yop proteins once they are secreted by yersiniae.

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    Derivative Assay:

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    Article Snippet: The stock (10mg/ml in PBS/BSA) solution of rabbit IgG, derived from goats, (diluted to 50 pg/ml ~ 50 μg/ml) was used for determining the standard curve. .. Then 100μl of anti-rabbit IgG-horseradish peroxidase (HRP) conjugate (Sigma, St.Louis, MO) was added to each well, and the solution was incubated for one hour on a plate orbital shaker.

    Immunohistochemistry:

    Article Title: Therapeutic effect of nanoliposomal PCSK9 vaccine in a mouse model of atherosclerosis
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    Concentration Assay:

    Article Title: Accelerated immunoassays based on magnetic particle dynamics in a rotating capillary tube with stationary magnetic field
    Article Snippet: The SPP particle based sandwich ELISA was performed with a particle concentration of 0.06 %wt . .. Then 100μl of anti-rabbit IgG-horseradish peroxidase (HRP) conjugate (Sigma, St.Louis, MO) was added to each well, and the solution was incubated for one hour on a plate orbital shaker.

    Cell Culture:

    Article Title: Evaluation of YadC protein delivered by live attenuated Salmonella as a vaccine against plague
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    Inhibition:

    Article Title: Molecular Cloning and Characterization of a Surface-Localized Adhesion Protein in Mycoplasma bovis Hubei-1 Strain
    Article Snippet: For the adhesion inhibition assay, rVpmaX was incubated with rabbit anti-rVpmaX serum at different dilutions before interacting with the coated proteins. .. After incubating for 1 h at 37°C, unbound proteins were removed by washing, and the interaction between proteins was evaluated by adding rabbit anti-rVpmaX serum and goat anti-rabbit IgG peroxidase conjugate (Sigma).

    Injection:

    Article Title: The Small Heat Shock Protein p26 Aids Development of Encysting Artemia Embryos, Prevents Spontaneous Diapause Termination and Protects against Stress
    Article Snippet: Detection of sHSPs by SDS Polyacrylamide Gel Electrophoresis and Immunoprobing of Western Blots To produce protein solutions for each lane of an SDS polyacrylamide gel 25 cysts from females injected with either p26 dsRNA, control solution or GFP dsRNA were collected by centrifugation for 1 min at 20 g in a microcentrifuge. .. The membranes were washed 3 times for 5 min in TBS containing 0.1% Tween 20 (TBS-Tween) and in 10 mM TRIS containing 1 M NaCl and 0.5% Tween 20, pH 7.4 (HST) 3 times for 5 min prior to incubation for 15 min in HRP-conjugated goat anti-rabbit IgG antibody (Sigma-Aldrich) diluted 1∶10000 in TBS.

    Article Title: Production of Native Bispecific Antibodies in Rabbits
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    Recombinant:

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    Article Title: Expression of recombinant Rhipicephalus (Boophilus) microplus, R. annulatus and R. decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris
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    Nucleic Acid Electrophoresis:

    Article Title: Evaluation of YadC protein delivered by live attenuated Salmonella as a vaccine against plague
    Article Snippet: Protein samples from bacterial pellets and culture supernatants were separated and analyzed by SDS-polyacrylamide gel electrophoresis with transfer to nitrocellulose membranes as previously described ( ). .. Then, horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G (IgG) (Sigma) was added in 1X PBS/3% skim milk.

    Article Title: Adenylate Cyclase Toxin from Bordetella pertussis Synergizes with Lipopolysaccharide To Promote Innate Interleukin-10 Production and Enhances the Induction of Th2 and Regulatory T Cells
    Article Snippet: Alternatively, proteins were transferred to a nitrocellulose membrane following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and probed with anti-His Tag antibodies (Santa Cruz Biotechnologies) or anti-CyaA antibodies (kind gift from Erik Hewlett). .. The bands were visualized by incubation with secondary anti-rabbit immunoglobulin G (IgG) horseradish peroxidase-conjugated antibodies (Sigma) and chemiluminescent supersignal detection system (Pierce).

    Flow Cytometry:

    Article Title: Expression of recombinant Rhipicephalus (Boophilus) microplus, R. annulatus and R. decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris
    Article Snippet: Protein samples of 2 μl were absorbed onto nitrocellulose membrane by gravity flow to perform the dot blot analysis. .. Membranes were then washed three times with TBS and incubated with an anti-rabbit IgG horseradish peroxidase (HRP) conjugate (Sigma-Aldrich) diluted 1:1000 in TBS.

    Labeling:

    Article Title: Vitronectins produced by human cirrhotic liver and CCl4‐treated rats differ in their glycosylation pattern and tissue remodeling activity
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    Size-exclusion Chromatography:

    Article Title: Production of Native Bispecific Antibodies in Rabbits
    Article Snippet: Paragraph title: Size-exclusion chromatography ... Furthermore, the IgG nature of the antibodies was determined by a HRP conjugated goat anti-rabbit IgG antibody (1∶20000, Sigma-Aldrich).

    Dot Blot:

    Article Title: An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles
    Article Snippet: Paragraph title: 2.5 SDS-PAGE, dot blot and Western blot ... Proteins were either visualized by Coomassie blue staining or electrophoretically transferred to polyvinlidene difluoride (PVDF) membrane (Amersham Pharmacia, Piscataway, NJ), and probed in succession with rabbit anti-rNV serum [ ] at dilution of 1:10,000 in 1% DMyPBS and goat anti-rabbit-IgG-horseradish peroxidase conjugate (Sigma-Aldrich, St. Louis, MO) diluted in 1:10,000 in 1% DMyPBS.

    Article Title: Expression of recombinant Rhipicephalus (Boophilus) microplus, R. annulatus and R. decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris
    Article Snippet: Paragraph title: SDS-PAGE, dot blot and Western blot analyses ... Membranes were then washed three times with TBS and incubated with an anti-rabbit IgG horseradish peroxidase (HRP) conjugate (Sigma-Aldrich) diluted 1:1000 in TBS.

    Immunodetection:

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    Article Snippet: .. For the immunodetection of the fusion protein, rabbit anti-His probe antibody (Santa Cruz Biotechnology, Santa Cruz, Calif.) and goat anti-rabbit immunoglobulin G (IgG)-horseradish peroxidase conjugate (Sigma, St. Louis, Mo.) were used. .. The light-emitting nonradioactive ECL kit (Amersham Life Sciences, Buckinghamshire, United Kingdom) was used for signal detection.

    Cell Adhesion Assay:

    Article Title: Molecular Cloning and Characterization of a Surface-Localized Adhesion Protein in Mycoplasma bovis Hubei-1 Strain
    Article Snippet: For the adhesion assay, the rVpmaX protein was serially diluted with PBST and applied to either coated or blank plates. .. After incubating for 1 h at 37°C, unbound proteins were removed by washing, and the interaction between proteins was evaluated by adding rabbit anti-rVpmaX serum and goat anti-rabbit IgG peroxidase conjugate (Sigma).

    Construct:

    Article Title: Expression of recombinant Rhipicephalus (Boophilus) microplus, R. annulatus and R. decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris
    Article Snippet: A standard curve was constructed with known amounts of recombinant Bm86 extracted from Gavac (Revetmex) and was used for semi-quantitative analysis in dot-blots. .. Membranes were then washed three times with TBS and incubated with an anti-rabbit IgG horseradish peroxidase (HRP) conjugate (Sigma-Aldrich) diluted 1:1000 in TBS.

    Polyacrylamide Gel Electrophoresis:

    Article Title: The Small Heat Shock Protein p26 Aids Development of Encysting Artemia Embryos, Prevents Spontaneous Diapause Termination and Protects against Stress
    Article Snippet: Paragraph title: Detection of sHSPs by SDS Polyacrylamide Gel Electrophoresis and Immunoprobing of Western Blots ... The membranes were washed 3 times for 5 min in TBS containing 0.1% Tween 20 (TBS-Tween) and in 10 mM TRIS containing 1 M NaCl and 0.5% Tween 20, pH 7.4 (HST) 3 times for 5 min prior to incubation for 15 min in HRP-conjugated goat anti-rabbit IgG antibody (Sigma-Aldrich) diluted 1∶10000 in TBS.

    Article Title: Display of Bacterial Lipase on the Escherichia coli Cell Surface by Using FadL as an Anchoring Motif and Use of the Enzyme in Enantioselective Biocatalysis
    Article Snippet: Whole-cell lysates and membrane fractions were analyzed by SDS-12% (wt/vol) PAGE. .. For the immunodetection of the fusion protein, rabbit anti-His probe antibody (Santa Cruz Biotechnology, Santa Cruz, Calif.) and goat anti-rabbit immunoglobulin G (IgG)-horseradish peroxidase conjugate (Sigma, St. Louis, Mo.) were used.

    Purification:

    Article Title: An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles
    Article Snippet: Plant protein crude extracts and insect cells purified rNV were denatured by boiling in SDS–PAGE sample buffer and separated on 4–20% gradient polyacrylamide gels. .. Proteins were either visualized by Coomassie blue staining or electrophoretically transferred to polyvinlidene difluoride (PVDF) membrane (Amersham Pharmacia, Piscataway, NJ), and probed in succession with rabbit anti-rNV serum [ ] at dilution of 1:10,000 in 1% DMyPBS and goat anti-rabbit-IgG-horseradish peroxidase conjugate (Sigma-Aldrich, St. Louis, MO) diluted in 1:10,000 in 1% DMyPBS.

    Article Title: Adenylate Cyclase Toxin from Bordetella pertussis Synergizes with Lipopolysaccharide To Promote Innate Interleukin-10 Production and Enhances the Induction of Th2 and Regulatory T Cells
    Article Snippet: Paragraph title: Purification of CyaA. ... The bands were visualized by incubation with secondary anti-rabbit immunoglobulin G (IgG) horseradish peroxidase-conjugated antibodies (Sigma) and chemiluminescent supersignal detection system (Pierce).

    Article Title: Vitronectins produced by human cirrhotic liver and CCl4‐treated rats differ in their glycosylation pattern and tissue remodeling activity
    Article Snippet: Rabbit anti‐VN IgGs were purchased from the Cosmo Bio Co., Ltd (Tokyo, Japan), and HRP‐conjugated goat anti‐rabbit IgG was purchased from Millipore (Temecula, CA, USA). .. P. velutina lectin (PVL) was purified from the fruiting bodies of P. velutina mushrooms collected in Japan , .

    SDS Page:

    Article Title: Evaluation of YadC protein delivered by live attenuated Salmonella as a vaccine against plague
    Article Snippet: Paragraph title: 2.4 SDS-PAGE and western blot analysis ... Then, horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin G (IgG) (Sigma) was added in 1X PBS/3% skim milk.

    Article Title: An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles
    Article Snippet: Paragraph title: 2.5 SDS-PAGE, dot blot and Western blot ... Proteins were either visualized by Coomassie blue staining or electrophoretically transferred to polyvinlidene difluoride (PVDF) membrane (Amersham Pharmacia, Piscataway, NJ), and probed in succession with rabbit anti-rNV serum [ ] at dilution of 1:10,000 in 1% DMyPBS and goat anti-rabbit-IgG-horseradish peroxidase conjugate (Sigma-Aldrich, St. Louis, MO) diluted in 1:10,000 in 1% DMyPBS.

    Article Title: LcrV of Yersinia pestis Enters Infected Eukaryotic Cells by a Virulence Plasmid-Independent Mechanism
    Article Snippet: Proteins from fractionated, infected HeLa cultures were resolved in 12% (wt/vol) polyacrylamide gels by SDS-PAGE ( ). .. Proteins were visualized by treatment of immunoblots with alkaline phosphatase- or horseradish peroxidase-conjugated goat anti-rabbit IgG (Sigma), followed by development with nitroblue tetrazolium–5-bromo-4-chloro-3-indolylphosphate (NBT-BCIP; GIBCO-BRL) or enhanced chemiluminescence (ECL) substrate (Pierce), respectively.

    Article Title: Expression of recombinant Rhipicephalus (Boophilus) microplus, R. annulatus and R. decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris
    Article Snippet: Paragraph title: SDS-PAGE, dot blot and Western blot analyses ... Membranes were then washed three times with TBS and incubated with an anti-rabbit IgG horseradish peroxidase (HRP) conjugate (Sigma-Aldrich) diluted 1:1000 in TBS.

    Electrophoresis:

    Article Title: The Small Heat Shock Protein p26 Aids Development of Encysting Artemia Embryos, Prevents Spontaneous Diapause Termination and Protects against Stress
    Article Snippet: Nitrocellulose membranes containing cyst proteins separated by electrophoresis were incubated in 5% Carnation low fat milk solution for 45 min at room temperature, then in antibody specific to p26 diluted 1∶10000 in 10 mM TRIS containing 140 mM NaCl, pH 7.4 (TBS) for 20 min. .. The membranes were washed 3 times for 5 min in TBS containing 0.1% Tween 20 (TBS-Tween) and in 10 mM TRIS containing 1 M NaCl and 0.5% Tween 20, pH 7.4 (HST) 3 times for 5 min prior to incubation for 15 min in HRP-conjugated goat anti-rabbit IgG antibody (Sigma-Aldrich) diluted 1∶10000 in TBS.

    Negative Control:

    Article Title: Expression of recombinant Rhipicephalus (Boophilus) microplus, R. annulatus and R. decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris
    Article Snippet: The supernatant of the GS115/Albumin strain (Invitrogen) grown under the same conditions was used as a negative control in both dot- and Western-blots. .. Membranes were then washed three times with TBS and incubated with an anti-rabbit IgG horseradish peroxidase (HRP) conjugate (Sigma-Aldrich) diluted 1:1000 in TBS.

    Binding Assay:

    Article Title: Vitronectins produced by human cirrhotic liver and CCl4‐treated rats differ in their glycosylation pattern and tissue remodeling activity
    Article Snippet: Materials Sheep anti‐human VN IgGs were purchased from the Binding Site Ltd (Birmingham, UK), and horseradish peroxidase (HRP)‐conjugated rabbit anti‐sheep IgGs were purchased from ICN Biomedicals, Inc. (Costa Mesa, CA, USA). .. Rabbit anti‐VN IgGs were purchased from the Cosmo Bio Co., Ltd (Tokyo, Japan), and HRP‐conjugated goat anti‐rabbit IgG was purchased from Millipore (Temecula, CA, USA).

    Protein Electrophoresis:

    Article Title: LcrV of Yersinia pestis Enters Infected Eukaryotic Cells by a Virulence Plasmid-Independent Mechanism
    Article Snippet: Paragraph title: Protein electrophoresis and immunoblot analysis. ... Proteins were visualized by treatment of immunoblots with alkaline phosphatase- or horseradish peroxidase-conjugated goat anti-rabbit IgG (Sigma), followed by development with nitroblue tetrazolium–5-bromo-4-chloro-3-indolylphosphate (NBT-BCIP; GIBCO-BRL) or enhanced chemiluminescence (ECL) substrate (Pierce), respectively.

    Staining:

    Article Title: The Small Heat Shock Protein p26 Aids Development of Encysting Artemia Embryos, Prevents Spontaneous Diapause Termination and Protects against Stress
    Article Snippet: Fifteen µl of each protein sample was resolved in 12.5% SDS polyacrylamide gels and either stained with Coomassie blue or transferred to nitrocellulose membranes (Bio-Rad, Mississauga, ON, Canada). .. The membranes were washed 3 times for 5 min in TBS containing 0.1% Tween 20 (TBS-Tween) and in 10 mM TRIS containing 1 M NaCl and 0.5% Tween 20, pH 7.4 (HST) 3 times for 5 min prior to incubation for 15 min in HRP-conjugated goat anti-rabbit IgG antibody (Sigma-Aldrich) diluted 1∶10000 in TBS.

    Article Title: Therapeutic effect of nanoliposomal PCSK9 vaccine in a mouse model of atherosclerosis
    Article Snippet: .. The samples were then rinsed in PBS and incubated with goat anti-rabbit HRP-conjugated IgG antibody (Sigma-Aldrich Inc., St. Louis, MO) at the dilution of 1:200 for additional 60 min. To counterstain the samples, the nuclei were stained with hematoxylin. .. The sections were dehydrated and mounted with Entellan (Merck KGaA, Darmstadt, Germany).

    Article Title: An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles
    Article Snippet: .. Proteins were either visualized by Coomassie blue staining or electrophoretically transferred to polyvinlidene difluoride (PVDF) membrane (Amersham Pharmacia, Piscataway, NJ), and probed in succession with rabbit anti-rNV serum [ ] at dilution of 1:10,000 in 1% DMyPBS and goat anti-rabbit-IgG-horseradish peroxidase conjugate (Sigma-Aldrich, St. Louis, MO) diluted in 1:10,000 in 1% DMyPBS. .. The membranes were developed by chemiluminescence using the ECL plus detection reagent (Amersham Pharmacia).

    Article Title: Adenylate Cyclase Toxin from Bordetella pertussis Synergizes with Lipopolysaccharide To Promote Innate Interleukin-10 Production and Enhances the Induction of Th2 and Regulatory T Cells
    Article Snippet: Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and visualized with Coomassie blue (GelCode Blue Stain Reagent; Pierce). .. The bands were visualized by incubation with secondary anti-rabbit immunoglobulin G (IgG) horseradish peroxidase-conjugated antibodies (Sigma) and chemiluminescent supersignal detection system (Pierce).

    other:

    Article Title: SUMO Modification of NZFP Mediates Transcriptional Repression through TBP Binding
    Article Snippet: Rabbit anti-SUMO-1 (Santa Cruz), mouse anti-Flag (Sigma-Aldrich), mouse anti-GFP (Clontech), mouse anti-HA (Sigma-Aldrich), mouse anti-Myc (Santa Cruz) antibodies, anti-mouse Cy3-conjugated IgG, horseradish peroxidase (HRP)-linked goat anti-mouse and anti-rabbit IgG antibodies (Sigma-Aldrich) were purchased commercially.

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  • 88
    Millipore rabbit monoclonal anti inos
    <t>NPY</t> inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on <t>iNOS</t> (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated
    Rabbit Monoclonal Anti Inos, supplied by Millipore, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Millipore goat anti rabbit igg fitc conjugated
    Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary <t>anti-IgG-HRP</t> antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary <t>anti-IgG-FITC</t> antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.
    Goat Anti Rabbit Igg Fitc Conjugated, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore anti bdnf
    Effects of the ethanol withdrawal procedure on the hyperpolarization-activated cyclic nucleotide-gated cation channel <t>(HCN1)</t> and brain-derived neurotrophic factor <t>(BDNF)</t> protein or gene level changes in the nucleus accumbens (NAc). (A) The expression of BDNF BDNF mRNA in the NAc. (B) The expression of BDNF-positive cells in the NAc. (C) Expression of BDNF positive cells in the NAc. (D) The number of BDNF-positive cells in the Hip. (E) The expression of HCN1 mRNA in the NAc. (F) The expression of HCN1-positive cells in the NAc. (G) Expression of HCN1 positive cells in the NAc. (H) The number of HCN1-positive cells in the NAc. The BDNF- and HCN1-positive cells in the NAc are represented by photomicrographs (400×). Data are mean ± standard error. * P
    Anti Bdnf, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    NPY inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on iNOS (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated

    Journal: The Journal of Biological Chemistry

    Article Title: Neuropeptide Y Modulation of Interleukin-1? (IL-1?)-induced Nitric Oxide Production in Microglia *

    doi: 10.1074/jbc.M110.164020

    Figure Lengend Snippet: NPY inhibits inducible nitric-oxide synthase expression. Microglial cells were treated with 1 μ m NPY and challenged with 100 ng/ml LPS for 6 h to assess the effect of NPY on iNOS (130 kDa) protein levels. A , NPY significantly inhibited LPS-stimulated

    Article Snippet: Antibodies used were as follows: rabbit polyclonal anti-NPY (1:1000) (Sigma); sheep polyclonal anti-Y1 R (1:1000) (AbD Serotec, Oxfordshire, UK); rabbit monoclonal anti-iNOS (1:250) (Millipore Corp., Bedford, MA); rat monoclonal anti-CD11b (1:1000) (AbD Serotec); rabbit monoclonal anti-NF-κB p65 (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) in 0.1% Triton X-100, 0.3% BSA solution; and Alexa Fluor 594 goat anti-rabbit, Alexa Fluor 594 donkey anti-sheep, Alexa Fluor 594 goat anti-rat, Alexa Fluor 488 donkey anti-rabbit, and Alexa Fluor 488 goat anti-rat (all 1:200 in PBS, from Molecular Probes, Eugene, OR).

    Techniques: Expressing

    NPY inhibits IL-1β-induced iNOS protein levels. A , confocal microscopy photomicrographs illustrate microglial cells treated with 1 μ m NPY and 1.5 ng/ml IL-1β for 6 h to assess the role of NPY and IL-1β in iNOS synthesis.

    Journal: The Journal of Biological Chemistry

    Article Title: Neuropeptide Y Modulation of Interleukin-1? (IL-1?)-induced Nitric Oxide Production in Microglia *

    doi: 10.1074/jbc.M110.164020

    Figure Lengend Snippet: NPY inhibits IL-1β-induced iNOS protein levels. A , confocal microscopy photomicrographs illustrate microglial cells treated with 1 μ m NPY and 1.5 ng/ml IL-1β for 6 h to assess the role of NPY and IL-1β in iNOS synthesis.

    Article Snippet: Antibodies used were as follows: rabbit polyclonal anti-NPY (1:1000) (Sigma); sheep polyclonal anti-Y1 R (1:1000) (AbD Serotec, Oxfordshire, UK); rabbit monoclonal anti-iNOS (1:250) (Millipore Corp., Bedford, MA); rat monoclonal anti-CD11b (1:1000) (AbD Serotec); rabbit monoclonal anti-NF-κB p65 (1:100) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) in 0.1% Triton X-100, 0.3% BSA solution; and Alexa Fluor 594 goat anti-rabbit, Alexa Fluor 594 donkey anti-sheep, Alexa Fluor 594 goat anti-rat, Alexa Fluor 488 donkey anti-rabbit, and Alexa Fluor 488 goat anti-rat (all 1:200 in PBS, from Molecular Probes, Eugene, OR).

    Techniques: Confocal Microscopy

    Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary anti-IgG-FITC antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.

    Journal: Alcoholism, clinical and experimental research

    Article Title: Effects of Alcohol on Histone Deacetylase 2 (HDAC2) and the Neuroprotective Role of Trichostatin A (TSA)

    doi: 10.1111/j.1530-0277.2011.01492.x

    Figure Lengend Snippet: Alcohol Induces HDAC2 Protein and this Effect is Inhibited by TSA After reaching confluency, SK-N-MC were pre-incubated with TSA for 2 hours, then treated with EtOH (0.1%) for 48 hours. In figure 3a, 10 µg of protein were analyzed using western blot with primary anti-HDAC2 and secondary anti-IgG-HRP antibodies. GAPDH was used as a loading control. Data presented show a representative blot indicating modulation of HDAC2 protein expression and a bar graph representing the mean ± SE of % densitometry values of HDAC2 protein levels (% control) of three independent experiments. # represents significance compared to control. * represents significance compared to EtOH treatment. For the flow cytometry experiments, 1 × 10 6 cells were fixed and permeabilized prior to intracellular staining with primary anti-HDAC2 and secondary anti-IgG-FITC antibody. Data presented in figure 3b show a representative histogram overlay of the gated cells. The bar graph represents the mean ± SE of % of gated cells expressing HDAC2. 10000 events were analyzed per sample. The gray and black histograms represent the unlabeled and secondary antibody controls respectively; the green histogram is the untreated control (~52%), blue represents EtOH (~69 %), purple represents TSA (~45%), and orange represents TSA + EtOH (~49%) treated cells. Data are representative of three independent experiments.

    Article Snippet: The HDAC2 protein was detected with the primary monoclonal antibody, rabbit anti-histone deacetylase 2 (Millipore) and secondary antibody, goat anti-rabbit IgG FITC-conjugated (Millipore).

    Techniques: Incubation, Western Blot, Expressing, Flow Cytometry, Cytometry, Staining

    Effects of the ethanol withdrawal procedure on the hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN1) and brain-derived neurotrophic factor (BDNF) protein or gene level changes in the nucleus accumbens (NAc). (A) The expression of BDNF BDNF mRNA in the NAc. (B) The expression of BDNF-positive cells in the NAc. (C) Expression of BDNF positive cells in the NAc. (D) The number of BDNF-positive cells in the Hip. (E) The expression of HCN1 mRNA in the NAc. (F) The expression of HCN1-positive cells in the NAc. (G) Expression of HCN1 positive cells in the NAc. (H) The number of HCN1-positive cells in the NAc. The BDNF- and HCN1-positive cells in the NAc are represented by photomicrographs (400×). Data are mean ± standard error. * P

    Journal: Frontiers in Psychiatry

    Article Title: Synaptic Ultrastructure Might Be Involved in HCN1-Related BDNF mRNA in Withdrawal-Anxiety After Ethanol Dependence

    doi: 10.3389/fpsyt.2018.00215

    Figure Lengend Snippet: Effects of the ethanol withdrawal procedure on the hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN1) and brain-derived neurotrophic factor (BDNF) protein or gene level changes in the nucleus accumbens (NAc). (A) The expression of BDNF BDNF mRNA in the NAc. (B) The expression of BDNF-positive cells in the NAc. (C) Expression of BDNF positive cells in the NAc. (D) The number of BDNF-positive cells in the Hip. (E) The expression of HCN1 mRNA in the NAc. (F) The expression of HCN1-positive cells in the NAc. (G) Expression of HCN1 positive cells in the NAc. (H) The number of HCN1-positive cells in the NAc. The BDNF- and HCN1-positive cells in the NAc are represented by photomicrographs (400×). Data are mean ± standard error. * P

    Article Snippet: The next day, the membrane was washed three times, 10 min each time, with PBS-T and incubated at room temperature with secondary antibodies, including anti-HCN1 [Peroxidase-Conjugated Affinipure Goat Anti-mouse IgG (H + L), 1:200] and anti-BDNF [Peroxidase-Conjugated Affinipure Goat Anti-Rabbit IgG (H + L), 1:200], and reacted with the Immobilon western chemiluminescent HRP substrate (WBKLS0500; Millipore, Bedford, MA, USA) for the color reaction.

    Techniques: Derivative Assay, Expressing

    Effects of the ethanol withdrawal procedure on the hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN1) and brain-derived neurotrophic factor (BDNF) protein or gene level changes in the hippocampus (Hip). (A) The expression of BDNF BDNF mRNA in the Hip. (B) The expression of BDNF-positive cells in the Hip. (C) Expression of BDNF positive cells in the Hip. (D) The number of BDNF-positive cells in the Hip. (E) The expression of HCN1 mRNA in the Hip. (F) The expression of HCN1-positive cells in the Hip. (G) Expression of HCN1 positive cells in Hip. (H) The number of HCN1-positive cells in the Hip. The BDNF- and HCN1-positive cells in the Hip are represented by photomicrographs (400×). Data are mean ± standard error. * P

    Journal: Frontiers in Psychiatry

    Article Title: Synaptic Ultrastructure Might Be Involved in HCN1-Related BDNF mRNA in Withdrawal-Anxiety After Ethanol Dependence

    doi: 10.3389/fpsyt.2018.00215

    Figure Lengend Snippet: Effects of the ethanol withdrawal procedure on the hyperpolarization-activated cyclic nucleotide-gated cation channel (HCN1) and brain-derived neurotrophic factor (BDNF) protein or gene level changes in the hippocampus (Hip). (A) The expression of BDNF BDNF mRNA in the Hip. (B) The expression of BDNF-positive cells in the Hip. (C) Expression of BDNF positive cells in the Hip. (D) The number of BDNF-positive cells in the Hip. (E) The expression of HCN1 mRNA in the Hip. (F) The expression of HCN1-positive cells in the Hip. (G) Expression of HCN1 positive cells in Hip. (H) The number of HCN1-positive cells in the Hip. The BDNF- and HCN1-positive cells in the Hip are represented by photomicrographs (400×). Data are mean ± standard error. * P

    Article Snippet: The next day, the membrane was washed three times, 10 min each time, with PBS-T and incubated at room temperature with secondary antibodies, including anti-HCN1 [Peroxidase-Conjugated Affinipure Goat Anti-mouse IgG (H + L), 1:200] and anti-BDNF [Peroxidase-Conjugated Affinipure Goat Anti-Rabbit IgG (H + L), 1:200], and reacted with the Immobilon western chemiluminescent HRP substrate (WBKLS0500; Millipore, Bedford, MA, USA) for the color reaction.

    Techniques: Derivative Assay, Expressing