biotinylated monoclonal anti rabbit igg  (Millipore)

 
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    Name:
    Anti Rabbit IgG gamma chain specific Biotin antibody
    Description:
    The product recognizes an epitope located on the gamma heavy chain of rabbit IgG In immunoblotting the antibody recognizes both native and denatured forms of rabbit IgG In ELISA the antibody is specific for rabbit IgG and shows no cross reactivity with rabbit IgA and IgM or human IgG IgA and IgM No cross reaction is observed with IgG from the following species bovine cat chicken dog goat guinea pig horse pig rat or sheep
    Catalog Number:
    B5283
    Price:
    None
    Applications:
    Monoclonal Anti-Rabbit IgG (γ-chain specific)-Biotin is suitable for ELISA at a working dilution of 1:150,000. It may be used for immunohistochemistry of formalin-fixed, paraffin-embedded sections at a dilution of 1:1000. The antibody was used in β-catenin ELISA at a dilution of 1:2000.Anti-Rabbit IgG (γ-chain specific)-Biotin antibody, Mouse monoclonal has been used in β-Catenin enzyme immunoassay. It has also been used in immunohistochemistry for examining histology of mammary glands.
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    Structured Review

    Millipore biotinylated monoclonal anti rabbit igg
    The product recognizes an epitope located on the gamma heavy chain of rabbit IgG In immunoblotting the antibody recognizes both native and denatured forms of rabbit IgG In ELISA the antibody is specific for rabbit IgG and shows no cross reactivity with rabbit IgA and IgM or human IgG IgA and IgM No cross reaction is observed with IgG from the following species bovine cat chicken dog goat guinea pig horse pig rat or sheep
    https://www.bioz.com/result/biotinylated monoclonal anti rabbit igg/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated monoclonal anti rabbit igg - by Bioz Stars, 2021-07
    93/100 stars

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    Incubation:

    Article Title: Plant expression, lyophilisation and storage of HBV medium and large surface antigens for a prototype oral vaccine formulation
    Article Snippet: .. All the following incubations with extracts or antibodies were conducted at room temperature for 45 min. After sample incubation, wells were incubated with 1:4,000 anti-HBsAg domain S-specific polyclonal rabbit antibody (B65811R, Meridian Life Science), then with 1:20,000 anti-rabbit IgG γ-specific biotin-conjugated mAb (Sigma), followed by 1:4,000 AP-conjugated ExtrAvidin® (Sigma). .. The reaction with pNPP substrate (Sigma) was developed at 25°C for 1 h and absorbance was measured at 405 nm using a Microplate Reader, Model 680 (Bio-Rad, USA).

    Article Title: Heterogeneity in heat shock response dynamics caused by translation fidelity decline and proteostasis collapse
    Article Snippet: Lastly, the membrane for identification of Tubulin, as control protein, was incubated in 5% milk powder in TBS-t buffer with anti-tubulin primary antibody (1:500, Sigma #T9026, Buchs, Switzerland)). .. All membranes with primary antibodies were incubated overnight at 4° C. Then, after through TBS washes, blots were probed with secondary antibodies: Tubulin and GFP with anti-mouse IgG antibody (1:2000) and HSP-16.2 with anti-rabbit IgG antibody (1:1500) in TBS-t suspensions with 5% milk powder for 1 h and 1.5 h respectively. .. Finally, the protein bands were visualized upon few minutes of incubation of the membranes with Bio-Rad Clarity Western ECL Substrate (Bio-Rad#1705061, Cressier, Switzerland).

    Article Title: FERM domain–containing protein 6 identifies a subpopulation of varicose nerve fibers in different vertebrate species
    Article Snippet: Samples were separated on a 4–12% Bis-Tris PAGE under reducing conditions and blotted onto PVDF membranes. .. Blots were incubated with an anti-FRMD6 antiserum raised against the C-terminus of the protein and as secondary antibody, a biotinylated mouse anti-rabbit monoclonal antibody (γ-chain-specific, Clone RG-96) (Sigma-Aldrich) was used. .. Bands were visualized by streptavidin peroxidase incubation and chemiluminescence detection.

    Article Title: Sequence determinants in the cathelicidin LL-37 that promote inflammation via presentation of RNA to scavenger receptors
    Article Snippet: For LL-34 PLA binding, LL-37 antibody (sc-166770; Santa Cruz Biotechnologies, 1:1000 dilution) was used. .. After washing with 1× PBS, the cells were further incubated with Plus and Minus oligonucleotide probe–conjugated secondary antibodies to biotin (1:1000 dilution) and CD36 (1:1000 dilution) for 60 min at 37 °C in a humidified chamber. .. Further hybridization, ligation, amplification, and detection of the PLA was performed according to the manufacturer's instructions (Duolink, catalog no.: DUO92101, Sigma–Aldrich).

    Article Title: Lifelong Expression of Apolipoprotein D in the Human Brainstem: Correlation with Reduced Age-Related Neurodegeneration
    Article Snippet: Blocked membranes were then incubated for 1 hour at room temperature with a primary antibody against human Apo D (1:10,000; a gift from Dr. Carlos López Otín, Departamento de Bioquímica y Biología Molecular, Universidad de Oviedo) [ , - ]. .. After 3 washes of 15 min each in PBS, membranes were incubated for 30 min at room temperature in biotinylated monoclonal anti-rabbit IgG (1:10,000; Sigma, B 5283), washed again as before, and then incubated with ExtrAvidin Alkaline Phosphatase (1:10,000; Sigma, E26366). .. After three washes of 15 min each in PBS, enzyme activity was measured by incubation in Sigma Fast BCIP/NBT solution (Sigma, B5655) for 2 hours at room temperature.

    BAC Assay:

    Article Title: Localization and Expression of Zonula Occludins-1 in the Rabbit Corneal Epithelium following Exposure to Benzalkonium Chloride
    Article Snippet: We found that exposure to BAC can quickly disrupt the tight junctions between superficial cells in the rabbit corneal epithelium. .. Antibodies and Reagent BAC was purchased from Sigma (St. Louis, MO); pentobarbital sodium from Abbott Laboratories (North Chicago, IL); mouse anti-rabbit ZO-1 antibody, mouse anti-rabbit ZO-2 antibody, mouse anti-rabbit claudin-1 and mouse anti-rabbit occludin from Zymed (Carlsbad, CA); rat monoclonal antibodies for Ki67 from DakoCytomation (MIB-1; Glostrup, Denmark); TUNEL apoptosis detection kit was purchased from Roche Diagnostics (Meylen, France); Alexa488-conjugated goat anti-mouse IgG, goat anti-mouse IgG and Alexa594-conjuagated mouse anti-rat IgG from Molecular Probes (Eugene, OR); 4,6-diamidino-2-phenylindole (DAPI) and bovine serum albumin (BSA) from Vector Laboratories (Burlingame CA); Dulbecco modified Eagle medium (DMEM) and Dispase II from Invitrogen Corp (Carlsbad, CA); mouse anti-rabbit β actin antibody from Sigma; Horseradish peroxidase-conjugated goat antibody to mouse immunoglobulin G (IgG), nitrocellulose membranes and an enhanced chemiluminescence (ECL) kit from GE Healthcare UK (Chalfont, UK). .. Animals and BAC Treatment A total of 36 male white New Zealand rabbits (purchased from Shanghai Medical Laboratory Animal Center, Shanghai, China) weighing between 1.5 and 2.0 kg were randomly assigned to three groups of 12 rabbits each.

    TUNEL Assay:

    Article Title: Localization and Expression of Zonula Occludins-1 in the Rabbit Corneal Epithelium following Exposure to Benzalkonium Chloride
    Article Snippet: We found that exposure to BAC can quickly disrupt the tight junctions between superficial cells in the rabbit corneal epithelium. .. Antibodies and Reagent BAC was purchased from Sigma (St. Louis, MO); pentobarbital sodium from Abbott Laboratories (North Chicago, IL); mouse anti-rabbit ZO-1 antibody, mouse anti-rabbit ZO-2 antibody, mouse anti-rabbit claudin-1 and mouse anti-rabbit occludin from Zymed (Carlsbad, CA); rat monoclonal antibodies for Ki67 from DakoCytomation (MIB-1; Glostrup, Denmark); TUNEL apoptosis detection kit was purchased from Roche Diagnostics (Meylen, France); Alexa488-conjugated goat anti-mouse IgG, goat anti-mouse IgG and Alexa594-conjuagated mouse anti-rat IgG from Molecular Probes (Eugene, OR); 4,6-diamidino-2-phenylindole (DAPI) and bovine serum albumin (BSA) from Vector Laboratories (Burlingame CA); Dulbecco modified Eagle medium (DMEM) and Dispase II from Invitrogen Corp (Carlsbad, CA); mouse anti-rabbit β actin antibody from Sigma; Horseradish peroxidase-conjugated goat antibody to mouse immunoglobulin G (IgG), nitrocellulose membranes and an enhanced chemiluminescence (ECL) kit from GE Healthcare UK (Chalfont, UK). .. Animals and BAC Treatment A total of 36 male white New Zealand rabbits (purchased from Shanghai Medical Laboratory Animal Center, Shanghai, China) weighing between 1.5 and 2.0 kg were randomly assigned to three groups of 12 rabbits each.

    Modification:

    Article Title: Localization and Expression of Zonula Occludins-1 in the Rabbit Corneal Epithelium following Exposure to Benzalkonium Chloride
    Article Snippet: We found that exposure to BAC can quickly disrupt the tight junctions between superficial cells in the rabbit corneal epithelium. .. Antibodies and Reagent BAC was purchased from Sigma (St. Louis, MO); pentobarbital sodium from Abbott Laboratories (North Chicago, IL); mouse anti-rabbit ZO-1 antibody, mouse anti-rabbit ZO-2 antibody, mouse anti-rabbit claudin-1 and mouse anti-rabbit occludin from Zymed (Carlsbad, CA); rat monoclonal antibodies for Ki67 from DakoCytomation (MIB-1; Glostrup, Denmark); TUNEL apoptosis detection kit was purchased from Roche Diagnostics (Meylen, France); Alexa488-conjugated goat anti-mouse IgG, goat anti-mouse IgG and Alexa594-conjuagated mouse anti-rat IgG from Molecular Probes (Eugene, OR); 4,6-diamidino-2-phenylindole (DAPI) and bovine serum albumin (BSA) from Vector Laboratories (Burlingame CA); Dulbecco modified Eagle medium (DMEM) and Dispase II from Invitrogen Corp (Carlsbad, CA); mouse anti-rabbit β actin antibody from Sigma; Horseradish peroxidase-conjugated goat antibody to mouse immunoglobulin G (IgG), nitrocellulose membranes and an enhanced chemiluminescence (ECL) kit from GE Healthcare UK (Chalfont, UK). .. Animals and BAC Treatment A total of 36 male white New Zealand rabbits (purchased from Shanghai Medical Laboratory Animal Center, Shanghai, China) weighing between 1.5 and 2.0 kg were randomly assigned to three groups of 12 rabbits each.

    other:

    Article Title: Proteasome Inhibition Induces Nuclear Translocation and Transcriptional Activation of the Dioxin Receptor in Mouse Embryo Primary Fibroblasts in the Absence of Xenobiotics
    Article Snippet: Rabbit antibody against mouse β-actin was obtained from Sigma.

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  • 97
    Millipore mouse neun
    Enzymatic and metabolic correction of AspA deficiency by systemically delivered rAAV-mediated gene therapy in CD mice . ( a ) Western blot of brain homogenates from WT (P90), Het (P90), and CD mice treated with PBS (P27) or rAAV hAspA (P90). Loading control: GAPDH. ( b ) <t>Aspartoacylase</t> activity in brain homogenates of the study groups. ( c ) Persistent vector genomes (by quantitative PCR) and aspartoacylase expression (by western blot) in P90 CD/rAAV9hAspA and CD/rAAV9 hAspAmiRBS . ( d ) Representative images of avidin-biotin-complex (ABC) stained brain sections. ( e ) Immunofluorescence images of brain sections from P90 WT and CD/rAAV9 for AspA (green) and neuronal marker, <t>NeuN</t> (red) and colocalization (yellow). Bars: 10 μm. ( f ) Representative MRS spectra of brain NAA levels in WT and CD/PBS mice at P26 ( n = 3 each) showing NAA peaks at 2.018 ppm. ( g ) Quantification of in vivo brain metabolite levels from the proton spectra as ratio to total creatine. Ins: inositol, tCho: total choline, tNAA: total NAA, tCr: total creatine, Glx: glutamate + glutamine. ( h ) Mass spectrometry of urine from P27 WT, Het and CD/PBS or CD/rAAV9 mice. WT, wild-type; Het, heterozygote; CD/PBS, CD/rAAV9, CD mice treated with PBS or rAAV9 respectively. CD, Canavan's disease; MRS, magnetic resonance spectroscopy; NAA, N-acetyl aspartic acid; NS, not significant; PBS, phosphate-buffered saline; rAAV, recombinant adeno-associated virus.
    Mouse Neun, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse neun/product/Millipore
    Average 97 stars, based on 1 article reviews
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    97
    Millipore mouse anti human hab18g cd147 monoclonal antibody mab hab18
    Confocal microscopic analysis of <t>HAb18G/CD147</t> and antagonistic peptide (AP)–9. A Severe acute respiratory syndrome coronavirus (SARS-CoV)–infected 293 cells stained with biotin–AP-9 and avidin-Cy3 (red) . The result showed that AP-9 was bound to the detected cells. The binding of AP-9 to the detected cells was partially blocked by HAb18G/CD147. B Immunofluorescence double-labeling method in SARS-CoV–infected 293 cells. Two kinds of fluorescence that indicated HAb18G/CD147 (green) and AP-9 (red) simultaneously presented in the cells, as observed by confocal imaging
    Mouse Anti Human Hab18g Cd147 Monoclonal Antibody Mab Hab18, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti human hab18g cd147 monoclonal antibody mab hab18/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti human hab18g cd147 monoclonal antibody mab hab18 - by Bioz Stars, 2021-07
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    93
    Millipore biotinylated mouse anti rabbit monoclonal antibody
    Lane 1: <t>biotinylated</t> molecular weight marker. Lane 2: SDS extract of AtT-20 whole cell lysate probed with the C-terminus-specific antibody. To show the specificity of the antibody, lane 2 was cut into two halves prior to incubation with the primary antibody (cutting line indicated by a punctate vertical line) and both parts were incubated in separate trays with different antisera. The left side of lane 2 was incubated as control with an antiserum against green fluorescent protein, while the right part of lane 2 was incubated with the FRMD6 C-terminus-specific antiserum. Both parts of lane 2 were finally put together again before chemiluminescence imaging. A single specific band at 70 kDa is apparent solely on the right part of lane 2 exposed to the FRMD6 C-terminus-specific antibody, while the left part shows no signal at the corresponding molecular weight. Lanes 3–7: FRMD6 immunoprecipitation in different tissues with the FRMD6 N-terminus-specific antiserum. Immunoprecipitates (IPs) were probed with the FRMD6 C-terminus-specific antibody as in lane 2. Lane 3: a homogenate of rat spinal cord displays no significant band. Lanes 4 and 5: homogenates from two different AtT-20 cell cultures with identical cell densities. A specific band at 70 kDa but with variable intensity is apparent. Lanes 6 and 7: NIH3T3 and MCF-7 cultures, respectively. Lane 8: a control is shown performed under identical conditions with a parallel MCF-7 culture, except that for immunoprecipitation, a rabbit antiserum raised against an unrelated protein (human SGSM3) was used. The entire blotting membrane area is displayed in ESM, Fig. S3 a
    Biotinylated Mouse Anti Rabbit Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/biotinylated mouse anti rabbit monoclonal antibody/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    biotinylated mouse anti rabbit monoclonal antibody - by Bioz Stars, 2021-07
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    97
    Millipore gfap
    Lane 1: <t>biotinylated</t> molecular weight marker. Lane 2: SDS extract of AtT-20 whole cell lysate probed with the C-terminus-specific antibody. To show the specificity of the antibody, lane 2 was cut into two halves prior to incubation with the primary antibody (cutting line indicated by a punctate vertical line) and both parts were incubated in separate trays with different antisera. The left side of lane 2 was incubated as control with an antiserum against green fluorescent protein, while the right part of lane 2 was incubated with the FRMD6 C-terminus-specific antiserum. Both parts of lane 2 were finally put together again before chemiluminescence imaging. A single specific band at 70 kDa is apparent solely on the right part of lane 2 exposed to the FRMD6 C-terminus-specific antibody, while the left part shows no signal at the corresponding molecular weight. Lanes 3–7: FRMD6 immunoprecipitation in different tissues with the FRMD6 N-terminus-specific antiserum. Immunoprecipitates (IPs) were probed with the FRMD6 C-terminus-specific antibody as in lane 2. Lane 3: a homogenate of rat spinal cord displays no significant band. Lanes 4 and 5: homogenates from two different AtT-20 cell cultures with identical cell densities. A specific band at 70 kDa but with variable intensity is apparent. Lanes 6 and 7: NIH3T3 and MCF-7 cultures, respectively. Lane 8: a control is shown performed under identical conditions with a parallel MCF-7 culture, except that for immunoprecipitation, a rabbit antiserum raised against an unrelated protein (human SGSM3) was used. The entire blotting membrane area is displayed in ESM, Fig. S3 a
    Gfap, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfap/product/Millipore
    Average 97 stars, based on 1 article reviews
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    gfap - by Bioz Stars, 2021-07
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    Enzymatic and metabolic correction of AspA deficiency by systemically delivered rAAV-mediated gene therapy in CD mice . ( a ) Western blot of brain homogenates from WT (P90), Het (P90), and CD mice treated with PBS (P27) or rAAV hAspA (P90). Loading control: GAPDH. ( b ) Aspartoacylase activity in brain homogenates of the study groups. ( c ) Persistent vector genomes (by quantitative PCR) and aspartoacylase expression (by western blot) in P90 CD/rAAV9hAspA and CD/rAAV9 hAspAmiRBS . ( d ) Representative images of avidin-biotin-complex (ABC) stained brain sections. ( e ) Immunofluorescence images of brain sections from P90 WT and CD/rAAV9 for AspA (green) and neuronal marker, NeuN (red) and colocalization (yellow). Bars: 10 μm. ( f ) Representative MRS spectra of brain NAA levels in WT and CD/PBS mice at P26 ( n = 3 each) showing NAA peaks at 2.018 ppm. ( g ) Quantification of in vivo brain metabolite levels from the proton spectra as ratio to total creatine. Ins: inositol, tCho: total choline, tNAA: total NAA, tCr: total creatine, Glx: glutamate + glutamine. ( h ) Mass spectrometry of urine from P27 WT, Het and CD/PBS or CD/rAAV9 mice. WT, wild-type; Het, heterozygote; CD/PBS, CD/rAAV9, CD mice treated with PBS or rAAV9 respectively. CD, Canavan's disease; MRS, magnetic resonance spectroscopy; NAA, N-acetyl aspartic acid; NS, not significant; PBS, phosphate-buffered saline; rAAV, recombinant adeno-associated virus.

    Journal: Molecular Therapy

    Article Title: A Single Intravenous rAAV Injection as Late as P20 Achieves Efficacious and Sustained CNS Gene Therapy in Canavan Mice

    doi: 10.1038/mt.2013.138

    Figure Lengend Snippet: Enzymatic and metabolic correction of AspA deficiency by systemically delivered rAAV-mediated gene therapy in CD mice . ( a ) Western blot of brain homogenates from WT (P90), Het (P90), and CD mice treated with PBS (P27) or rAAV hAspA (P90). Loading control: GAPDH. ( b ) Aspartoacylase activity in brain homogenates of the study groups. ( c ) Persistent vector genomes (by quantitative PCR) and aspartoacylase expression (by western blot) in P90 CD/rAAV9hAspA and CD/rAAV9 hAspAmiRBS . ( d ) Representative images of avidin-biotin-complex (ABC) stained brain sections. ( e ) Immunofluorescence images of brain sections from P90 WT and CD/rAAV9 for AspA (green) and neuronal marker, NeuN (red) and colocalization (yellow). Bars: 10 μm. ( f ) Representative MRS spectra of brain NAA levels in WT and CD/PBS mice at P26 ( n = 3 each) showing NAA peaks at 2.018 ppm. ( g ) Quantification of in vivo brain metabolite levels from the proton spectra as ratio to total creatine. Ins: inositol, tCho: total choline, tNAA: total NAA, tCr: total creatine, Glx: glutamate + glutamine. ( h ) Mass spectrometry of urine from P27 WT, Het and CD/PBS or CD/rAAV9 mice. WT, wild-type; Het, heterozygote; CD/PBS, CD/rAAV9, CD mice treated with PBS or rAAV9 respectively. CD, Canavan's disease; MRS, magnetic resonance spectroscopy; NAA, N-acetyl aspartic acid; NS, not significant; PBS, phosphate-buffered saline; rAAV, recombinant adeno-associated virus.

    Article Snippet: The primary antibodies used were as follows: mouse monoclonal antibodies to human aspartoacylase (Abmart, Shanghai, China), rabbit aspartoacylase (provided by Dr Aryan Namboodiri); mouse NeuN (MAB-377, clone A60; Millipore, Billerica, MA); rabbit NeuN (ABN-78; Millipore) and appropriate secondary antibodies from Invitrogen.

    Techniques: Mouse Assay, Western Blot, Activity Assay, Plasmid Preparation, Real-time Polymerase Chain Reaction, Expressing, Avidin-Biotin Assay, Staining, Immunofluorescence, Marker, In Vivo, Mass Spectrometry, Spectroscopy, Recombinant

    Confocal microscopic analysis of HAb18G/CD147 and antagonistic peptide (AP)–9. A Severe acute respiratory syndrome coronavirus (SARS-CoV)–infected 293 cells stained with biotin–AP-9 and avidin-Cy3 (red) . The result showed that AP-9 was bound to the detected cells. The binding of AP-9 to the detected cells was partially blocked by HAb18G/CD147. B Immunofluorescence double-labeling method in SARS-CoV–infected 293 cells. Two kinds of fluorescence that indicated HAb18G/CD147 (green) and AP-9 (red) simultaneously presented in the cells, as observed by confocal imaging

    Journal: The Journal of Infectious Diseases

    Article Title: Function of HAb18G/CD147 in Invasion of Host Cells by Severe Acute Respiratory Syndrome Coronavirus

    doi: 10.1086/427811

    Figure Lengend Snippet: Confocal microscopic analysis of HAb18G/CD147 and antagonistic peptide (AP)–9. A Severe acute respiratory syndrome coronavirus (SARS-CoV)–infected 293 cells stained with biotin–AP-9 and avidin-Cy3 (red) . The result showed that AP-9 was bound to the detected cells. The binding of AP-9 to the detected cells was partially blocked by HAb18G/CD147. B Immunofluorescence double-labeling method in SARS-CoV–infected 293 cells. Two kinds of fluorescence that indicated HAb18G/CD147 (green) and AP-9 (red) simultaneously presented in the cells, as observed by confocal imaging

    Article Snippet: The mixture was then added to the gel, which was coupled with 200 μg of mouse anti–human HAb18G/CD147 monoclonal antibody (MAb) HAb18 (prepared in our lab) [ ], for incubation with gentle end-over-end mixing for 4 h. After the bound proteins were eluted, aliquots of the eluent were analyzed by a Western blot developed with rabbit anti–human CyPA antibody (diluted 1:2000; Calbiochem-Novabiochem) and horseradish peroxidase (HRP)–conjugated goat anti–rabbit IgG (diluted 1:5000; Amersham Pharmacia).

    Techniques: Infection, Staining, Avidin-Biotin Assay, Binding Assay, Immunofluorescence, Labeling, Fluorescence, Imaging

    Subcellular localization of cyclophilin A (CyPA), HAb18G/CD147, and antagonistic peptide (AP)–9 on severe acute respiratory syndrome coronavirus (SARS-CoV)–infected 293 cells. A Gold particles representing CyPA (arrow) located on the virus surface in a cluster or around the virus. B Gold particles representing CD147 (arrow) located on the infected cell surface, across the membrane, or on the unit membrane in cytoplasm (especially on the membrane of endoplasmic reticulum) in cluster. C Gold colloid double labeling. Gold colloid particles of 2 different sizes that indicated AP-9 (20 nm) and HAb18G/CD147 (10 nm), respectively, were located on the same site (arrow)

    Journal: The Journal of Infectious Diseases

    Article Title: Function of HAb18G/CD147 in Invasion of Host Cells by Severe Acute Respiratory Syndrome Coronavirus

    doi: 10.1086/427811

    Figure Lengend Snippet: Subcellular localization of cyclophilin A (CyPA), HAb18G/CD147, and antagonistic peptide (AP)–9 on severe acute respiratory syndrome coronavirus (SARS-CoV)–infected 293 cells. A Gold particles representing CyPA (arrow) located on the virus surface in a cluster or around the virus. B Gold particles representing CD147 (arrow) located on the infected cell surface, across the membrane, or on the unit membrane in cytoplasm (especially on the membrane of endoplasmic reticulum) in cluster. C Gold colloid double labeling. Gold colloid particles of 2 different sizes that indicated AP-9 (20 nm) and HAb18G/CD147 (10 nm), respectively, were located on the same site (arrow)

    Article Snippet: The mixture was then added to the gel, which was coupled with 200 μg of mouse anti–human HAb18G/CD147 monoclonal antibody (MAb) HAb18 (prepared in our lab) [ ], for incubation with gentle end-over-end mixing for 4 h. After the bound proteins were eluted, aliquots of the eluent were analyzed by a Western blot developed with rabbit anti–human CyPA antibody (diluted 1:2000; Calbiochem-Novabiochem) and horseradish peroxidase (HRP)–conjugated goat anti–rabbit IgG (diluted 1:5000; Amersham Pharmacia).

    Techniques: Infection, Labeling

    Analysis of protein-protein interaction. A Coimmunoprecipitation (Co-IP) analysis–revealed interaction between HAb18G/CD147 and cyclophilin A (CyPA). Blank, blank control; eluate, coimmunoprecipitated CyPA in the eluate; standard, 2.5 μg of purchased CyPA. B Co-IP analysis–revealed interaction between CyPA and severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid (N) protein. Blank, blank control; eluate 1, 2, and 3, coimmunoprecipitated N protein in orderly eluting; standard, 5 μg of N protein expressed in Escherichia coli. C Binding kinetics of SARS-CoV N protein to CyPA, determined by surface plasmon resonance analysis. N protein, at different concentrations, could bind to CyPA. The lines with different colors are the binding kinetics curves of N protein to CyPA, at different concentrations: 40, 32, 24, 16, and 8 nmol/L. RU, resonance unit

    Journal: The Journal of Infectious Diseases

    Article Title: Function of HAb18G/CD147 in Invasion of Host Cells by Severe Acute Respiratory Syndrome Coronavirus

    doi: 10.1086/427811

    Figure Lengend Snippet: Analysis of protein-protein interaction. A Coimmunoprecipitation (Co-IP) analysis–revealed interaction between HAb18G/CD147 and cyclophilin A (CyPA). Blank, blank control; eluate, coimmunoprecipitated CyPA in the eluate; standard, 2.5 μg of purchased CyPA. B Co-IP analysis–revealed interaction between CyPA and severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid (N) protein. Blank, blank control; eluate 1, 2, and 3, coimmunoprecipitated N protein in orderly eluting; standard, 5 μg of N protein expressed in Escherichia coli. C Binding kinetics of SARS-CoV N protein to CyPA, determined by surface plasmon resonance analysis. N protein, at different concentrations, could bind to CyPA. The lines with different colors are the binding kinetics curves of N protein to CyPA, at different concentrations: 40, 32, 24, 16, and 8 nmol/L. RU, resonance unit

    Article Snippet: The mixture was then added to the gel, which was coupled with 200 μg of mouse anti–human HAb18G/CD147 monoclonal antibody (MAb) HAb18 (prepared in our lab) [ ], for incubation with gentle end-over-end mixing for 4 h. After the bound proteins were eluted, aliquots of the eluent were analyzed by a Western blot developed with rabbit anti–human CyPA antibody (diluted 1:2000; Calbiochem-Novabiochem) and horseradish peroxidase (HRP)–conjugated goat anti–rabbit IgG (diluted 1:5000; Amersham Pharmacia).

    Techniques: Co-Immunoprecipitation Assay, Binding Assay, SPR Assay

    Flow-cytometric analysis of HAb18G/CD147 and antagonistic peptide (AP)–9. A Expression of HAb18G/CD147 on 293 cells, analyzed by use of fluorescein isothiocyanate (FITC)–conjugated anti-CD147 antibody. B Binding of AP-9 to 293 cells, analyzed by use of biotin–AP-9 and avidin-FITC

    Journal: The Journal of Infectious Diseases

    Article Title: Function of HAb18G/CD147 in Invasion of Host Cells by Severe Acute Respiratory Syndrome Coronavirus

    doi: 10.1086/427811

    Figure Lengend Snippet: Flow-cytometric analysis of HAb18G/CD147 and antagonistic peptide (AP)–9. A Expression of HAb18G/CD147 on 293 cells, analyzed by use of fluorescein isothiocyanate (FITC)–conjugated anti-CD147 antibody. B Binding of AP-9 to 293 cells, analyzed by use of biotin–AP-9 and avidin-FITC

    Article Snippet: The mixture was then added to the gel, which was coupled with 200 μg of mouse anti–human HAb18G/CD147 monoclonal antibody (MAb) HAb18 (prepared in our lab) [ ], for incubation with gentle end-over-end mixing for 4 h. After the bound proteins were eluted, aliquots of the eluent were analyzed by a Western blot developed with rabbit anti–human CyPA antibody (diluted 1:2000; Calbiochem-Novabiochem) and horseradish peroxidase (HRP)–conjugated goat anti–rabbit IgG (diluted 1:5000; Amersham Pharmacia).

    Techniques: Expressing, Binding Assay, Avidin-Biotin Assay

    Lane 1: biotinylated molecular weight marker. Lane 2: SDS extract of AtT-20 whole cell lysate probed with the C-terminus-specific antibody. To show the specificity of the antibody, lane 2 was cut into two halves prior to incubation with the primary antibody (cutting line indicated by a punctate vertical line) and both parts were incubated in separate trays with different antisera. The left side of lane 2 was incubated as control with an antiserum against green fluorescent protein, while the right part of lane 2 was incubated with the FRMD6 C-terminus-specific antiserum. Both parts of lane 2 were finally put together again before chemiluminescence imaging. A single specific band at 70 kDa is apparent solely on the right part of lane 2 exposed to the FRMD6 C-terminus-specific antibody, while the left part shows no signal at the corresponding molecular weight. Lanes 3–7: FRMD6 immunoprecipitation in different tissues with the FRMD6 N-terminus-specific antiserum. Immunoprecipitates (IPs) were probed with the FRMD6 C-terminus-specific antibody as in lane 2. Lane 3: a homogenate of rat spinal cord displays no significant band. Lanes 4 and 5: homogenates from two different AtT-20 cell cultures with identical cell densities. A specific band at 70 kDa but with variable intensity is apparent. Lanes 6 and 7: NIH3T3 and MCF-7 cultures, respectively. Lane 8: a control is shown performed under identical conditions with a parallel MCF-7 culture, except that for immunoprecipitation, a rabbit antiserum raised against an unrelated protein (human SGSM3) was used. The entire blotting membrane area is displayed in ESM, Fig. S3 a

    Journal: Cell and Tissue Research

    Article Title: FERM domain–containing protein 6 identifies a subpopulation of varicose nerve fibers in different vertebrate species

    doi: 10.1007/s00441-020-03189-7

    Figure Lengend Snippet: Lane 1: biotinylated molecular weight marker. Lane 2: SDS extract of AtT-20 whole cell lysate probed with the C-terminus-specific antibody. To show the specificity of the antibody, lane 2 was cut into two halves prior to incubation with the primary antibody (cutting line indicated by a punctate vertical line) and both parts were incubated in separate trays with different antisera. The left side of lane 2 was incubated as control with an antiserum against green fluorescent protein, while the right part of lane 2 was incubated with the FRMD6 C-terminus-specific antiserum. Both parts of lane 2 were finally put together again before chemiluminescence imaging. A single specific band at 70 kDa is apparent solely on the right part of lane 2 exposed to the FRMD6 C-terminus-specific antibody, while the left part shows no signal at the corresponding molecular weight. Lanes 3–7: FRMD6 immunoprecipitation in different tissues with the FRMD6 N-terminus-specific antiserum. Immunoprecipitates (IPs) were probed with the FRMD6 C-terminus-specific antibody as in lane 2. Lane 3: a homogenate of rat spinal cord displays no significant band. Lanes 4 and 5: homogenates from two different AtT-20 cell cultures with identical cell densities. A specific band at 70 kDa but with variable intensity is apparent. Lanes 6 and 7: NIH3T3 and MCF-7 cultures, respectively. Lane 8: a control is shown performed under identical conditions with a parallel MCF-7 culture, except that for immunoprecipitation, a rabbit antiserum raised against an unrelated protein (human SGSM3) was used. The entire blotting membrane area is displayed in ESM, Fig. S3 a

    Article Snippet: Blots were incubated with an anti-FRMD6 antiserum raised against the C-terminus of the protein and as secondary antibody, a biotinylated mouse anti-rabbit monoclonal antibody (γ-chain-specific, Clone RG-96) (Sigma-Aldrich) was used.

    Techniques: Molecular Weight, Marker, Incubation, Imaging, Immunoprecipitation