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GE Healthcare hrp conjugated goat anti rabbit igg
Hrp Conjugated Goat Anti Rabbit Igg, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 94/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/hrp conjugated goat anti rabbit igg/product/GE Healthcare
Average 94 stars, based on 45 article reviews
Price from $9.99 to $1999.99
hrp conjugated goat anti rabbit igg - by Bioz Stars, 2020-04
94/100 stars

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Related Articles

Whole Genome Amplification:

Article Title: Tunicamycin-induced ER stress in breast cancer cells neither expresses GRP78 on the surface nor secretes it into the media
Article Snippet: WGA (Texas-Red conjugated) and Concanavalin A (FITC-conjugated) were from EY Laboratories (San Mateo, CA). .. HRP-conjugated goat anti-rabbit IgG, HRP-conjugated rabbit anti-mouse IgG, HRP-conjugated streptavidin, and ECL chemiluminescence detection kit were from GE Healthcare (Piscataway, NJ).

Bradford Assay:

Article Title: Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate. Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate
Article Snippet: Total protein extracted was quantified using Bradford assay (dye from Bio‐Rad, USA), and different concentrations of bovine serum albumin (BSA) (Sigma‐Aldrich) solubilized in urea buffer were used as standards. .. After three washes with PBS, the blot was incubated with 1:10,000 dilution of HRP‐conjugated goat anti‐rabbit IgG for 10 min, washed three times, and developed using the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare, UK).

Electrophoresis:

Article Title: Tunicamycin-induced ER stress in breast cancer cells neither expresses GRP78 on the surface nor secretes it into the media
Article Snippet: HRP-conjugated goat anti-rabbit IgG, HRP-conjugated rabbit anti-mouse IgG, HRP-conjugated streptavidin, and ECL chemiluminescence detection kit were from GE Healthcare (Piscataway, NJ). .. Biotinylated protein molecular weight markers and all electrophoresis reagents were obtained from Bio-Rad Laboratories (Hercules, CA).

Incubation:

Article Title: CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers
Article Snippet: .. Membranes were incubated with CD44 antibody (Clone 156-3C11) for 2 hours at RT followed by horse-radish-peroxidase conjugated goat anti-rabbit IgG (GE Healthcare, Chalfont, St. Giles, UK) for 1 hour at RT in 5% dry-milk with TBST. .. Visualization was performed using ECL Plus (GE Healthcare).

Article Title: THE DANGER SIGNAL, EXTRACELLULAR ATP, IS A SENSOR FOR AN AIRBORNE ALLERGEN AND TRIGGERS IL-33 RELEASE AND INNATE TH2-TYPE RESPONSES 1
Article Snippet: After protein transfer, the nitrocellulose membrane was cut, blocked in milk buffer, and incubated with normal rabbit IgG or rabbit anti-human IL-33 (1 μg/ml). .. HRP-conjugated goat anti-rabbit IgG was used for detection, and the blot was developed using an ECL kit (Amersham Pharmacia Biotech),followed by exposure to Biomax® film (Kodak).

Article Title: Transport of estradiol-17?-glucuronide, estrone-3-sulfate and taurocholate across the endoplasmic reticulum membrane: evidence for different transport systems
Article Snippet: Membrane fractions were probed for Cyp3a2 (Enzo Lifesciences CR3320, Lausen, Switzerland; 1:7000 dilution), Sec61α (Thermo Scientific PA3-014, Waltham, Massachusetts; dilution 1:7000) or PDI (Thermo scientific MA3-019, Waltham, Massachusetts; dilution 1:20 000 or 1:100 000) by either incubation at 4 °C overnight (Sec61α) or at room temperature for 2 h (Cyp3a2; PDI). .. Either HRP-conjugated goat anti rabbit IgG (GE Healthcare RPN4301, Little Chalfont, United Kingdom; for Sec61α and Cyp3a2; dilution 1:30 000) or sheep anti mouse IgG (GE Healthcare NA931VS, Little Chalfont, United Kingdom; dilution 1:3000) was used as secondary antibody.

Article Title: Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate. Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate
Article Snippet: .. After three washes with PBS, the blot was incubated with 1:10,000 dilution of HRP‐conjugated goat anti‐rabbit IgG for 10 min, washed three times, and developed using the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare, UK). .. The chemiluminescence was visualized under FUSION‐SL Chemiluminescence System (Vilber Lourmat, France).

Article Title: Sensing and signaling of oxidative stress in chloroplasts by inactivation of the SAL1 phosphoadenosine phosphatase
Article Snippet: .. After three washes with PBST, the blot was incubated with 1:10,000 dilution of HRP-conjugated goat anti-rabbit IgG for 10 min. After a further three washes with PBST, the membrane was developed with the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare). .. Gel bands were manually excised for identification by mass spectrometry.

Expressing:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: Paragraph title: Evaluation of GFP/β3 -Endonexin Expression in CHO Cells ... After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL).

BIA-KA:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: After clarification of the lysate in a microfuge, protein concentration was determined with a bicinchoninic acid reagent (BCA; Pierce Chemical Co. ). .. After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL).

Western Blot:

Article Title: GLUT4 and UBC9 Protein Expression Is Reduced in Muscle from Type 2 Diabetic Patients with Severe Insulin Resistance
Article Snippet: Paragraph title: Western Blot analysis ... HRP-conjugated goat anti-rabbit IgG (#NA934, GE healthcare) was used as secondary antibody.

Article Title: CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers
Article Snippet: Paragraph title: Western blot ... Membranes were incubated with CD44 antibody (Clone 156-3C11) for 2 hours at RT followed by horse-radish-peroxidase conjugated goat anti-rabbit IgG (GE Healthcare, Chalfont, St. Giles, UK) for 1 hour at RT in 5% dry-milk with TBST.

Article Title: Transport of estradiol-17?-glucuronide, estrone-3-sulfate and taurocholate across the endoplasmic reticulum membrane: evidence for different transport systems
Article Snippet: Paragraph title: Western blotting ... Either HRP-conjugated goat anti rabbit IgG (GE Healthcare RPN4301, Little Chalfont, United Kingdom; for Sec61α and Cyp3a2; dilution 1:30 000) or sheep anti mouse IgG (GE Healthcare NA931VS, Little Chalfont, United Kingdom; dilution 1:3000) was used as secondary antibody.

Article Title: PRESYNAPTIC CALCIUM CHANNELS AND ?3-INTEGRINS ARE COMPLEXED WITH SYNAPTIC CLEFT LAMININS, CYTOSKELETAL ELEMENTS AND ACTIVE ZONE COMPONENTS
Article Snippet: .. HRP conjugated goat anti-rabbit IgG, and HRP conjugated goat anti-mouse IgG used for Western blots were purchased from GE Healthcare. ..

Article Title: Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate. Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate
Article Snippet: .. After three washes with PBS, the blot was incubated with 1:10,000 dilution of HRP‐conjugated goat anti‐rabbit IgG for 10 min, washed three times, and developed using the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare, UK). .. The chemiluminescence was visualized under FUSION‐SL Chemiluminescence System (Vilber Lourmat, France).

Article Title: Sensing and signaling of oxidative stress in chloroplasts by inactivation of the SAL1 phosphoadenosine phosphatase
Article Snippet: .. After three washes with PBST, the blot was incubated with 1:10,000 dilution of HRP-conjugated goat anti-rabbit IgG for 10 min. After a further three washes with PBST, the membrane was developed with the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare). .. Gel bands were manually excised for identification by mass spectrometry.

SDS-Gel:

Article Title: GLUT4 and UBC9 Protein Expression Is Reduced in Muscle from Type 2 Diabetic Patients with Severe Insulin Resistance
Article Snippet: In short, protein was separated on a 4–12% Bis-Tris SDS gel (CriterionTM XT, Bio-Rad, CA, USA), transferred to a PVDF membrane, and immunoblotted. β-actin (Abcam, #ab82227-50) was used as a control for equal loading. .. HRP-conjugated goat anti-rabbit IgG (#NA934, GE healthcare) was used as secondary antibody.

Transfection:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: 48 h after transfection, the cells were lysed for 30 min at 4°C in a lysis buffer containing 1% Triton X-100, 0.9% NaCl, 1 mM CaCl2 , 50 mM Tris, pH 7.2, and protease inhibitors (100 U/ml aprotinin, 0.5 mM leupeptin, 4 mM Pefabloc) ( ). .. After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL).

Protease Inhibitor:

Article Title: CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers
Article Snippet: Western blot Cells were lysed in RIPA buffer containing protease inhibitor (Roche, Basel, Swizerland) and mixed with 2× NuPage sample buffer. .. Membranes were incubated with CD44 antibody (Clone 156-3C11) for 2 hours at RT followed by horse-radish-peroxidase conjugated goat anti-rabbit IgG (GE Healthcare, Chalfont, St. Giles, UK) for 1 hour at RT in 5% dry-milk with TBST.

Cell Culture:

Article Title: Tunicamycin-induced ER stress in breast cancer cells neither expresses GRP78 on the surface nor secretes it into the media
Article Snippet: HRP-conjugated goat anti-rabbit IgG, HRP-conjugated rabbit anti-mouse IgG, HRP-conjugated streptavidin, and ECL chemiluminescence detection kit were from GE Healthcare (Piscataway, NJ). .. Cell culture plastic wares were from Sarstedt (Newton, NC).

Generated:

Article Title: GLUT4 and UBC9 Protein Expression Is Reduced in Muscle from Type 2 Diabetic Patients with Severe Insulin Resistance
Article Snippet: Antibodies against GLUT4 were generated as previously described and commercially available antibodies against: AS160 (Cell Signalling, #2447), TBC1D1 (Abcam, #ab56191-100), UBC9 (Abcam, #ab75854), and GLUT1 (Millipore, #07-1401) were used. .. HRP-conjugated goat anti-rabbit IgG (#NA934, GE healthcare) was used as secondary antibody.

Article Title: PRESYNAPTIC CALCIUM CHANNELS AND ?3-INTEGRINS ARE COMPLEXED WITH SYNAPTIC CLEFT LAMININS, CYTOSKELETAL ELEMENTS AND ACTIVE ZONE COMPONENTS
Article Snippet: These fusion proteins were generated from bassoon clone pGBK-RB25 obtained from Drs. W. Altrock and E. Gundelfinger (Magdeburg, Gemany). .. HRP conjugated goat anti-rabbit IgG, and HRP conjugated goat anti-mouse IgG used for Western blots were purchased from GE Healthcare.

Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates
Article Snippet: Antibody binding and competition curves were generated by subtracting background controls from assay signals. .. The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

Inhibition:

Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates
Article Snippet: The assay established the inhibition of plate-immobilized IVIg F(ab) fragments (200 ng/well) binding to 8 μg/mL of solution-phase PFs by serially diluted (6 nM to 6 μM) human and murine pAb competitors. .. The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

Protein Concentration:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: After clarification of the lysate in a microfuge, protein concentration was determined with a bicinchoninic acid reagent (BCA; Pierce Chemical Co. ). .. After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL).

Affinity Purification:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: .. After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL). .. To study the binding of GFP/β3 -endonexin to the β3 integrin cytoplasmic tail, the 47–amino acid β3 tail was expressed in bacteria with a (His)6 tag at its amino terminus (pET His Tag System; Novagen, Inc., Madison, WI), and then immobilized on a nickel-agarose matrix.

Binding Assay:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL). .. To study the binding of GFP/β3 -endonexin to the β3 integrin cytoplasmic tail, the 47–amino acid β3 tail was expressed in bacteria with a (His)6 tag at its amino terminus (pET His Tag System; Novagen, Inc., Madison, WI), and then immobilized on a nickel-agarose matrix.

Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates
Article Snippet: Paragraph title: Antibody binding to amyloidogenic conformers-ELISAs ... The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

Molecular Weight:

Article Title: Tunicamycin-induced ER stress in breast cancer cells neither expresses GRP78 on the surface nor secretes it into the media
Article Snippet: HRP-conjugated goat anti-rabbit IgG, HRP-conjugated rabbit anti-mouse IgG, HRP-conjugated streptavidin, and ECL chemiluminescence detection kit were from GE Healthcare (Piscataway, NJ). .. Biotinylated protein molecular weight markers and all electrophoresis reagents were obtained from Bio-Rad Laboratories (Hercules, CA).

MTT Assay:

Article Title: Tunicamycin-induced ER stress in breast cancer cells neither expresses GRP78 on the surface nor secretes it into the media
Article Snippet: Dimethyl sulfoxide, nystatin, cell dissociation solution, tunicamycin and methylthiazolyl diphenyl tetrazolium bromide (MTT) were obtained from Sigma-Aldrich (St. Louis, MO). .. HRP-conjugated goat anti-rabbit IgG, HRP-conjugated rabbit anti-mouse IgG, HRP-conjugated streptavidin, and ECL chemiluminescence detection kit were from GE Healthcare (Piscataway, NJ).

Fluorescence:

Article Title: PRESYNAPTIC CALCIUM CHANNELS AND ?3-INTEGRINS ARE COMPLEXED WITH SYNAPTIC CLEFT LAMININS, CYTOSKELETAL ELEMENTS AND ACTIVE ZONE COMPONENTS
Article Snippet: Secondary antibodies and toxins used for fluorescence microscopy were purchased from Invitrogen and are as follows: Alexa 488 and 568 conjugated donkey anti-rabbit IgG, Alexa 488 and 568 conjugated donkey anti-mouse IgG, and Alexa 647 conjugated α-bungarotoxin. .. HRP conjugated goat anti-rabbit IgG, and HRP conjugated goat anti-mouse IgG used for Western blots were purchased from GE Healthcare.

Isolation:

Article Title: Transport of estradiol-17?-glucuronide, estrone-3-sulfate and taurocholate across the endoplasmic reticulum membrane: evidence for different transport systems
Article Snippet: Either HRP-conjugated goat anti rabbit IgG (GE Healthcare RPN4301, Little Chalfont, United Kingdom; for Sec61α and Cyp3a2; dilution 1:30 000) or sheep anti mouse IgG (GE Healthcare NA931VS, Little Chalfont, United Kingdom; dilution 1:3000) was used as secondary antibody. .. Western blot analyses for Cyp3a2 and Sec61α were performed for each SER and RER isolation used in this study.

Clarification Assay:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: After clarification of the lysate in a microfuge, protein concentration was determined with a bicinchoninic acid reagent (BCA; Pierce Chemical Co. ). .. After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL).

Immunohistochemistry:

Article Title: Cyclopamine tartrate, a modulator of hedgehog signaling and mitochondrial respiration, effectively arrests lung tumor growth and progression
Article Snippet: Paragraph title: Immunohistochemistry (IHC) ... Antibodies for CA9 (100-417), HRP-conjugated goat anti-mouse IgG (NB7539) and for Hexokinase II (PA5-29326), HRP-conjugated goat anti-rabbit IgG (#31460) were purchased from Novus and Thermo Fisher Scientific, respectively.

Microscopy:

Article Title: PRESYNAPTIC CALCIUM CHANNELS AND ?3-INTEGRINS ARE COMPLEXED WITH SYNAPTIC CLEFT LAMININS, CYTOSKELETAL ELEMENTS AND ACTIVE ZONE COMPONENTS
Article Snippet: Secondary antibodies and toxins used for fluorescence microscopy were purchased from Invitrogen and are as follows: Alexa 488 and 568 conjugated donkey anti-rabbit IgG, Alexa 488 and 568 conjugated donkey anti-mouse IgG, and Alexa 647 conjugated α-bungarotoxin. .. HRP conjugated goat anti-rabbit IgG, and HRP conjugated goat anti-mouse IgG used for Western blots were purchased from GE Healthcare.

Positron Emission Tomography:

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL). .. To study the binding of GFP/β3 -endonexin to the β3 integrin cytoplasmic tail, the 47–amino acid β3 tail was expressed in bacteria with a (His)6 tag at its amino terminus (pET His Tag System; Novagen, Inc., Madison, WI), and then immobilized on a nickel-agarose matrix.

SDS Page:

Article Title: Transport of estradiol-17?-glucuronide, estrone-3-sulfate and taurocholate across the endoplasmic reticulum membrane: evidence for different transport systems
Article Snippet: 2.4 Western blotting Proteins were separated by SDS-PAGE using 7.5% separating gels followed by blotting on nitrocellulose membranes (GE Healthcare, Little Chalfont, United Kingdom) with standard procedures. .. Either HRP-conjugated goat anti rabbit IgG (GE Healthcare RPN4301, Little Chalfont, United Kingdom; for Sec61α and Cyp3a2; dilution 1:30 000) or sheep anti mouse IgG (GE Healthcare NA931VS, Little Chalfont, United Kingdom; dilution 1:3000) was used as secondary antibody.

Article Title: Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate. Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate
Article Snippet: In brief, 5 μg of leaf total protein extract and 5 ng of recombinant SAL1 (rSAL1) used as a positive antibody specificity control were resolved on 4%–12% (w/v) SDS‐PAGE (NuPAGE; Invitrogen), electrotransferred to a PVDF membrane and probed with a 1:1,000 dilution of polyclonal antibodies raised against rSAL1 (Wilson et al., ) for 10 min using the SNAP i.d. system (Millipore, USA). .. After three washes with PBS, the blot was incubated with 1:10,000 dilution of HRP‐conjugated goat anti‐rabbit IgG for 10 min, washed three times, and developed using the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare, UK).

Article Title: Sensing and signaling of oxidative stress in chloroplasts by inactivation of the SAL1 phosphoadenosine phosphatase
Article Snippet: Five micrograms total Arabidopsis leaf protein were resolved by SDS/PAGE as described above, in absence of additional redox agents, then electrotransferred to a PVDF membrane and probed with a 1:1,000 dilution of polyclonal antibodies raised against AtSAL1 (Catalog Number AS07 256; Agrisera) as described previously ( ). .. After three washes with PBST, the blot was incubated with 1:10,000 dilution of HRP-conjugated goat anti-rabbit IgG for 10 min. After a further three washes with PBST, the membrane was developed with the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare).

Software:

Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates
Article Snippet: The resultant graphs were fit using a standard 3-parameter sigmoid (logistic) function (SigmaPlot 2000, version 6; Systat Software, Chicago, IL). .. The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

Enzyme-linked Immunosorbent Assay:

Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates
Article Snippet: The ability of human and murine pAbs to compete with each other for the same or similar binding sites on amyloidogenic conformers was determined using a hybrid capture/competition ELISA. .. The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

Recombinant:

Article Title: Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate. Development of strategies for genetic manipulation and fine‐tuning of a chloroplast retrograde signal 3′‐phosphoadenosine 5′‐phosphate
Article Snippet: In brief, 5 μg of leaf total protein extract and 5 ng of recombinant SAL1 (rSAL1) used as a positive antibody specificity control were resolved on 4%–12% (w/v) SDS‐PAGE (NuPAGE; Invitrogen), electrotransferred to a PVDF membrane and probed with a 1:1,000 dilution of polyclonal antibodies raised against rSAL1 (Wilson et al., ) for 10 min using the SNAP i.d. system (Millipore, USA). .. After three washes with PBS, the blot was incubated with 1:10,000 dilution of HRP‐conjugated goat anti‐rabbit IgG for 10 min, washed three times, and developed using the Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare, UK).

In Situ:

Article Title: Cyclopamine tartrate, a modulator of hedgehog signaling and mitochondrial respiration, effectively arrests lung tumor growth and progression
Article Snippet: Antibodies for CA9 (100-417), HRP-conjugated goat anti-mouse IgG (NB7539) and for Hexokinase II (PA5-29326), HRP-conjugated goat anti-rabbit IgG (#31460) were purchased from Novus and Thermo Fisher Scientific, respectively. .. The detection of DNA fragmentation in tumor tissues was carried out using the TumorTACSTM In Situ Apoptosis Detection Kit, following the manufacturer’s protocol.

Concentration Assay:

Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates
Article Snippet: The concentration of antibody that gave half-maximal binding, EC50 , and the concentration of competitor that gave half maximal inhibition, IC50 , were determined from the fitted curves. .. The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

Lysis:

Article Title: THE DANGER SIGNAL, EXTRACELLULAR ATP, IS A SENSOR FOR AN AIRBORNE ALLERGEN AND TRIGGERS IL-33 RELEASE AND INNATE TH2-TYPE RESPONSES 1
Article Snippet: NHBE cells (1×107 cells/ml) were lysed with RIPA lysis buffer (Santa Cruz). .. HRP-conjugated goat anti-rabbit IgG was used for detection, and the blot was developed using an ECL kit (Amersham Pharmacia Biotech),followed by exposure to Biomax® film (Kodak).

Article Title: Affinity Modulation of Platelet Integrin ?IIb?3 by ?3-Endonexin, a Selective Binding Partner of the ?3 Integrin Cytoplasmic Tail
Article Snippet: 48 h after transfection, the cells were lysed for 30 min at 4°C in a lysis buffer containing 1% Triton X-100, 0.9% NaCl, 1 mM CaCl2 , 50 mM Tris, pH 7.2, and protease inhibitors (100 U/ml aprotinin, 0.5 mM leupeptin, 4 mM Pefabloc) ( ). .. After the addition of affinity-purified, HRP-conjugated goat anti–rabbit IgG, the blots were developed for 0.1–1 min using the enhanced chemiluminescence reaction (ECL; Amersham Corp. , Arlington Heights, IL).

other:

Article Title: Potent Anti-Tumor Effect Generated by a Novel Human Papillomavirus (HPV) Antagonist Peptide Reactivating the pRb/E2F Pathway
Article Snippet: HRP-coupled mouse anti-M13 antibody, HRP-conjugated goat anti-mouse IgG and HRP-conjugated goat anti-rabbit IgG were supplied by Amersham Biosciences (Piscataway, NJ, USA) and KPL (Gaithersbug, MD, USA), respectively.

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  • 88
    GE Healthcare horse radish peroxidase conjugated goat anti rabbit igg
    A human mAb generated against a non-amyloid target binds aggregated <t>Aβ.</t> ( A ) Left panel: SEC chromatograms for ~15 mg/mL of mAb Avastin (anti-VEGF) and IVIg. SEC was carried out using a Superdex 200 Increase 10/300 GL column (GE Healthcare) equilibrated in PBS, pH 7.4. Right panel: Antibody binding curves against plate-immobilized PFs for unfractionated (Unfrac) IVIg, and for Avastin used unfractionated or as SEC-isolated monomers and dimers. ( B ) The top Western blots show immunoprecipitation (IP) of synthetic Aβ conformers (monomers (Mon), dimers, and PFs) by 100 μg/mL of Avastin and IVIg, and by 200 μg/mL of a pan-Aβ reactive polyclonal antibody, AW8. The blots were probed for Aβ using an Aβ N-terminal reactive mAb, 6E10 (Signet Laboratories). The lower Western blots show 20 μg/mL mAb Avastin's ability to IP 5 μg/mL of Aβ dimers and PFs in the presence of a 5-molar excess (with respect to Avastin) of a N-terminal 165-amino acid fragment of its immunogen VEGF (VEGF-165). Control IP experiments (Ctls) were carried out using 5 μg/mL mAb 6E10 and a mixture of Aβ dimers and PFs, or with 20 μg/mL Avastin and 1 μg/mL VEGF-165. The blots were probed for Aβ and VEGF-165 using mAb 6E10 and Avastin, respectively. In IPs carried out in the absence of VEGF-165, cross-reactivity of the secondary antibody, goat anti-human <t>IgG</t> (heavy and light, Jackson Immunoresearch Laboratories Inc), with Avastin’s Ig light chain caused a faint band that migrated near VEGF-165. ( C ) Avastin IgG conformers binding curves against plate-immobilized PFs in the presence or absence of a 1:10 dilution of IgG-depleted normal human sera. ( D ) Bar charts for solution-phase PF's, Aβ monomers, and non-amyloid native and aggregated molecule's inhibition of Avastin monomers an dimers binding to plate-immobilized PFs. Competition studies were carried out using 0.1 mg/mL competitors and concentrations of Avastin conformers that were equivalent to their EC 50 values for PFs: 500 nM IgG Mon, and 200 nM IgG dimer. Each competition curve was carried out in duplicate, and bars represent the standard error.
    Horse Radish Peroxidase Conjugated Goat Anti Rabbit Igg, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/horse radish peroxidase conjugated goat anti rabbit igg/product/GE Healthcare
    Average 88 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    horse radish peroxidase conjugated goat anti rabbit igg - by Bioz Stars, 2020-04
    88/100 stars
      Buy from Supplier

    86
    GE Healthcare secondary rabbit igg anti goat antibody conjugated to hrp
    Expression and characterization of TAP-tagged BRC motif in T. brucei (A) Map of the pLEW79FLR1TAP construct used for the in vivo doxycycline-inducible expression of a TAP-tagged single BRC motif (44 amino acids) with a 42 amino acid N-terminal flank. The recombinant protein (∼30 kDa) also has domains from calmodulin binding peptide (CBP, 26 amino acids), recognition site for TEV protease (TEV, 9 amino acids) and <t>IgG-binding</t> domains of Protein A (ProtA, 116 amino acids). Evaluation of the in vivo expression of the BRC motif-containing aptamer mRNA by RT-PCR (B) and corresponding protein by Western blot analysis (C). Since the wild-type procyclic T brucei cells (29-13) have tetracycline repressor protein (TetRep), the expression of the aptamer protein was inducible with doxycycline (Dox). RT-PCR products were generated with primers specific to TbBRCA2 BRC repeat. β-actin was used as a loading control in this analysis. Western Blot analysis was performed using a <t>HRP-conjugated</t> rabbit anti-goat IgG antibody. α-tubulin and the tet repressor were used as loading controls in this analysis. (D) Effect of expression of the BRC aptamer on the growth rate of procyclic forms of T. brucei . The controls included wild-type cells (WT 29-13) alone and wild-type cells transfected with the empty vector (Vector only). The other control is the recombinant cells grown in medium without doxycycline (No Dox cont). Results are mean ± SEM (n=12).
    Secondary Rabbit Igg Anti Goat Antibody Conjugated To Hrp, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/secondary rabbit igg anti goat antibody conjugated to hrp/product/GE Healthcare
    Average 86 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    secondary rabbit igg anti goat antibody conjugated to hrp - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    87
    GE Healthcare horseradish peroxidase hrp conjugated goat anti rabbit immunoglobulin g
    Localization of SERPINE2 in the human uterus . Longitudinal sections of the early secretory phase uterus (n = 5) on the slide were incubated with anti-mouse SERPINE2 antiserum and then treated with biotin-conjugated goat-anti-rabbit <t>IgG</t> and <t>HRP-conjugated</t> streptavidin (brown). For contrast, specimens were further stained with hematoxylin (blue). The representative picture is shown. Magnified pictures of the luminal epithelium (A), glandular epithelium (B), and myometrium (C) are shown. Bar = 50 μm. bv, blood vessel; ge, glandular epithelium; le, luminal epithelium; m, muscle; s, stroma.
    Horseradish Peroxidase Hrp Conjugated Goat Anti Rabbit Immunoglobulin G, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 87/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A human mAb generated against a non-amyloid target binds aggregated Aβ. ( A ) Left panel: SEC chromatograms for ~15 mg/mL of mAb Avastin (anti-VEGF) and IVIg. SEC was carried out using a Superdex 200 Increase 10/300 GL column (GE Healthcare) equilibrated in PBS, pH 7.4. Right panel: Antibody binding curves against plate-immobilized PFs for unfractionated (Unfrac) IVIg, and for Avastin used unfractionated or as SEC-isolated monomers and dimers. ( B ) The top Western blots show immunoprecipitation (IP) of synthetic Aβ conformers (monomers (Mon), dimers, and PFs) by 100 μg/mL of Avastin and IVIg, and by 200 μg/mL of a pan-Aβ reactive polyclonal antibody, AW8. The blots were probed for Aβ using an Aβ N-terminal reactive mAb, 6E10 (Signet Laboratories). The lower Western blots show 20 μg/mL mAb Avastin's ability to IP 5 μg/mL of Aβ dimers and PFs in the presence of a 5-molar excess (with respect to Avastin) of a N-terminal 165-amino acid fragment of its immunogen VEGF (VEGF-165). Control IP experiments (Ctls) were carried out using 5 μg/mL mAb 6E10 and a mixture of Aβ dimers and PFs, or with 20 μg/mL Avastin and 1 μg/mL VEGF-165. The blots were probed for Aβ and VEGF-165 using mAb 6E10 and Avastin, respectively. In IPs carried out in the absence of VEGF-165, cross-reactivity of the secondary antibody, goat anti-human IgG (heavy and light, Jackson Immunoresearch Laboratories Inc), with Avastin’s Ig light chain caused a faint band that migrated near VEGF-165. ( C ) Avastin IgG conformers binding curves against plate-immobilized PFs in the presence or absence of a 1:10 dilution of IgG-depleted normal human sera. ( D ) Bar charts for solution-phase PF's, Aβ monomers, and non-amyloid native and aggregated molecule's inhibition of Avastin monomers an dimers binding to plate-immobilized PFs. Competition studies were carried out using 0.1 mg/mL competitors and concentrations of Avastin conformers that were equivalent to their EC 50 values for PFs: 500 nM IgG Mon, and 200 nM IgG dimer. Each competition curve was carried out in duplicate, and bars represent the standard error.

    Journal: PLoS ONE

    Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates

    doi: 10.1371/journal.pone.0137344

    Figure Lengend Snippet: A human mAb generated against a non-amyloid target binds aggregated Aβ. ( A ) Left panel: SEC chromatograms for ~15 mg/mL of mAb Avastin (anti-VEGF) and IVIg. SEC was carried out using a Superdex 200 Increase 10/300 GL column (GE Healthcare) equilibrated in PBS, pH 7.4. Right panel: Antibody binding curves against plate-immobilized PFs for unfractionated (Unfrac) IVIg, and for Avastin used unfractionated or as SEC-isolated monomers and dimers. ( B ) The top Western blots show immunoprecipitation (IP) of synthetic Aβ conformers (monomers (Mon), dimers, and PFs) by 100 μg/mL of Avastin and IVIg, and by 200 μg/mL of a pan-Aβ reactive polyclonal antibody, AW8. The blots were probed for Aβ using an Aβ N-terminal reactive mAb, 6E10 (Signet Laboratories). The lower Western blots show 20 μg/mL mAb Avastin's ability to IP 5 μg/mL of Aβ dimers and PFs in the presence of a 5-molar excess (with respect to Avastin) of a N-terminal 165-amino acid fragment of its immunogen VEGF (VEGF-165). Control IP experiments (Ctls) were carried out using 5 μg/mL mAb 6E10 and a mixture of Aβ dimers and PFs, or with 20 μg/mL Avastin and 1 μg/mL VEGF-165. The blots were probed for Aβ and VEGF-165 using mAb 6E10 and Avastin, respectively. In IPs carried out in the absence of VEGF-165, cross-reactivity of the secondary antibody, goat anti-human IgG (heavy and light, Jackson Immunoresearch Laboratories Inc), with Avastin’s Ig light chain caused a faint band that migrated near VEGF-165. ( C ) Avastin IgG conformers binding curves against plate-immobilized PFs in the presence or absence of a 1:10 dilution of IgG-depleted normal human sera. ( D ) Bar charts for solution-phase PF's, Aβ monomers, and non-amyloid native and aggregated molecule's inhibition of Avastin monomers an dimers binding to plate-immobilized PFs. Competition studies were carried out using 0.1 mg/mL competitors and concentrations of Avastin conformers that were equivalent to their EC 50 values for PFs: 500 nM IgG Mon, and 200 nM IgG dimer. Each competition curve was carried out in duplicate, and bars represent the standard error.

    Article Snippet: The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

    Techniques: Generated, Size-exclusion Chromatography, Binding Assay, Isolation, Western Blot, Immunoprecipitation, Inhibition

    PAb aggregates have diverse avidities for Aβ. ( A ) Left panel: SEC chromatograms for untreated IVIg and for supernatants of ~4 mg/mL of SEC-isolated monomeric IVIg in PBS, pH 7.4, which was heated at 71°C until light scattering at A 400nm was 0.6 or 3.9, respectively. Right panel: Antibody binding curves against plate-immobilized PFs for untreated IVIg, Aβ-isolated IVIg IgGs, and for IgG supernatants (sup) and PBS resuspendend pellets (pellet) of heat-treated IVIg. (B ) Left panel: SEC chromatograms for ~4 mg/mL of untreated IVIg and for IVIg that was buffer exchanged at room temperature from gentle elution buffer (Pierce), pH 6.6, into PBS, pH 7.4. Right panel: Antibody binding curves against plate-immobilized PFs for untreated IVIg, buffered exchanged IVIg that was used unfractionated or as SEC-isolated monomers, and for Aβ-isolated IVIg IgGs.

    Journal: PLoS ONE

    Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates

    doi: 10.1371/journal.pone.0137344

    Figure Lengend Snippet: PAb aggregates have diverse avidities for Aβ. ( A ) Left panel: SEC chromatograms for untreated IVIg and for supernatants of ~4 mg/mL of SEC-isolated monomeric IVIg in PBS, pH 7.4, which was heated at 71°C until light scattering at A 400nm was 0.6 or 3.9, respectively. Right panel: Antibody binding curves against plate-immobilized PFs for untreated IVIg, Aβ-isolated IVIg IgGs, and for IgG supernatants (sup) and PBS resuspendend pellets (pellet) of heat-treated IVIg. (B ) Left panel: SEC chromatograms for ~4 mg/mL of untreated IVIg and for IVIg that was buffer exchanged at room temperature from gentle elution buffer (Pierce), pH 6.6, into PBS, pH 7.4. Right panel: Antibody binding curves against plate-immobilized PFs for untreated IVIg, buffered exchanged IVIg that was used unfractionated or as SEC-isolated monomers, and for Aβ-isolated IVIg IgGs.

    Article Snippet: The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

    Techniques: Size-exclusion Chromatography, Isolation, Binding Assay

    IVIg and Protein A-purified human and murine pAbs have similar anti-amyloid activities. ( A ) IgG binding curves against plate-immobilized PFs for IVIg and protein A-purified human (Hu) and murine (Mu) pAbs from pooled normal plasmas. Antibody binding curves are also shown for pAbs present in or dosed back into plasma. ( B ) Hybrid capture/competition ELISA curves for pAb's and IVIg's dose-dependent inhibition of PFs binding by plate-immobilized IVIg F(ab') fragments. The assay was carried out using 8 μg/ml solution-phase PFs. ( C ) Antibody binding curves for Hu and Mu pAb's nM cross-reactivity with plate-immobilized Aβ and TTR fibrils. ( D ) Left panel: Competition curves for solution-phase PF's and Aβ monomer's (Mon) inhibition of pAbs and IVIg binding to plate-immobilized PFs. Right panel: SAgg's and native TTR's (Nat) inhibition of pAbs and IVIg binding to plate-immobilized TTR fibrils. Competition studies were carried out using IgG concentrations, ~500 nM, which were equivalent to their EC 50 values for binding to PFs or TTR fibrils. Each binding or competition curve was carried out in duplicate, and bars represent the standard errors.

    Journal: PLoS ONE

    Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates

    doi: 10.1371/journal.pone.0137344

    Figure Lengend Snippet: IVIg and Protein A-purified human and murine pAbs have similar anti-amyloid activities. ( A ) IgG binding curves against plate-immobilized PFs for IVIg and protein A-purified human (Hu) and murine (Mu) pAbs from pooled normal plasmas. Antibody binding curves are also shown for pAbs present in or dosed back into plasma. ( B ) Hybrid capture/competition ELISA curves for pAb's and IVIg's dose-dependent inhibition of PFs binding by plate-immobilized IVIg F(ab') fragments. The assay was carried out using 8 μg/ml solution-phase PFs. ( C ) Antibody binding curves for Hu and Mu pAb's nM cross-reactivity with plate-immobilized Aβ and TTR fibrils. ( D ) Left panel: Competition curves for solution-phase PF's and Aβ monomer's (Mon) inhibition of pAbs and IVIg binding to plate-immobilized PFs. Right panel: SAgg's and native TTR's (Nat) inhibition of pAbs and IVIg binding to plate-immobilized TTR fibrils. Competition studies were carried out using IgG concentrations, ~500 nM, which were equivalent to their EC 50 values for binding to PFs or TTR fibrils. Each binding or competition curve was carried out in duplicate, and bars represent the standard errors.

    Article Snippet: The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

    Techniques: Purification, Binding Assay, Enzyme-linked Immunosorbent Assay, Inhibition

    IVIg IgG conformers retain binding to Aβ in the presence of normal human sera and non-amyloid molecules. ( A ) Antibody binding curves against plate-immobilized PFs for SEC-isolated IVIg monomers (SEC Mon), dimers (SEC Dimer), and for Aβ-isolated IVIg IgGs with or without a 1:10 dilution of IgG-depleted normal human sera. Preparations of Aβ-isolated IVIg IgGs contained HMW, dimeric, and monomeric species (see Fig 3A ). ( B ) Bar charts for solution-phase PFs, monomeric Aβ and for non-amyloid molecules inhibition of IVIg IgG conformers binding to plate-immobilized PFs. Non-amyloid molecules were chosen based on their: 1) Abundance in vivo (extracellular matrix and elastin fibrils), 2) Association with polyreactive autoantibodies (DNA), 3) High hydrophobicity (maize protein zein), and 4) Non-amyloid aggregate state [amorphous aggregated carboxymethylated ovalbumin (CM-Oval)]. Competition studies were carried out using 0.1 mg/mL competitors and concentrations of IgG conformers that were equivalent to their EC 50 values for PFs: 400 nM IgG Monomers; 50 nM IgG dimers, and 20 nM Aβ-isolated IgGs. Each competition curve was carried out in duplicate, and bars represent the standard error. ( C ) IVIg IgG conformer binding curves against PFs and non-amyloid molecules, murine extracellular matrix gel (ECM) and human DNA.

    Journal: PLoS ONE

    Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates

    doi: 10.1371/journal.pone.0137344

    Figure Lengend Snippet: IVIg IgG conformers retain binding to Aβ in the presence of normal human sera and non-amyloid molecules. ( A ) Antibody binding curves against plate-immobilized PFs for SEC-isolated IVIg monomers (SEC Mon), dimers (SEC Dimer), and for Aβ-isolated IVIg IgGs with or without a 1:10 dilution of IgG-depleted normal human sera. Preparations of Aβ-isolated IVIg IgGs contained HMW, dimeric, and monomeric species (see Fig 3A ). ( B ) Bar charts for solution-phase PFs, monomeric Aβ and for non-amyloid molecules inhibition of IVIg IgG conformers binding to plate-immobilized PFs. Non-amyloid molecules were chosen based on their: 1) Abundance in vivo (extracellular matrix and elastin fibrils), 2) Association with polyreactive autoantibodies (DNA), 3) High hydrophobicity (maize protein zein), and 4) Non-amyloid aggregate state [amorphous aggregated carboxymethylated ovalbumin (CM-Oval)]. Competition studies were carried out using 0.1 mg/mL competitors and concentrations of IgG conformers that were equivalent to their EC 50 values for PFs: 400 nM IgG Monomers; 50 nM IgG dimers, and 20 nM Aβ-isolated IgGs. Each competition curve was carried out in duplicate, and bars represent the standard error. ( C ) IVIg IgG conformer binding curves against PFs and non-amyloid molecules, murine extracellular matrix gel (ECM) and human DNA.

    Article Snippet: The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

    Techniques: Binding Assay, Size-exclusion Chromatography, Isolation, Inhibition, In Vivo

    Aβ-isolated but not heat-induced Avastin aggregates have enhanced avidity for PFs. ( A ) Left panel: SEC chromatograms for 0.3 mg/mL of Aβ-isolated Avastin IgGs, untreated Avastin, and for the antibody diluted into elution buffer (0.1 M glycine, pH 2.7) that was used to elute Aβ-bound Avastin IgGs. SEC was carried out using a Superdex 200 increase 10/300 GL column (GE Healthcare) that was equilibrated with PBS, pH 7.4. Right panel: Antibody binding curves against PFs for unfractionated IVIg and Avastin, and for Aβ-isolated Avastin IgGs. ( B ) Left panel: SEC chromatograms for ~5 mg/mL of unfractionated Avastin in PBS, pH 7.4, and for IgG conformers contained in supernatant of 71°C heated Avastin monomers (A 400nm 0.5 sup) in PBS, pH 7.4. Right panel: Antibody binding curves against PFs for soluble (A 400nm 0.5 sup) and insoluble (A 400nm 0.5 pellet) IgG conformers of heat-treated Avastin monomers, and for untreated Avastin and IVIg.

    Journal: PLoS ONE

    Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates

    doi: 10.1371/journal.pone.0137344

    Figure Lengend Snippet: Aβ-isolated but not heat-induced Avastin aggregates have enhanced avidity for PFs. ( A ) Left panel: SEC chromatograms for 0.3 mg/mL of Aβ-isolated Avastin IgGs, untreated Avastin, and for the antibody diluted into elution buffer (0.1 M glycine, pH 2.7) that was used to elute Aβ-bound Avastin IgGs. SEC was carried out using a Superdex 200 increase 10/300 GL column (GE Healthcare) that was equilibrated with PBS, pH 7.4. Right panel: Antibody binding curves against PFs for unfractionated IVIg and Avastin, and for Aβ-isolated Avastin IgGs. ( B ) Left panel: SEC chromatograms for ~5 mg/mL of unfractionated Avastin in PBS, pH 7.4, and for IgG conformers contained in supernatant of 71°C heated Avastin monomers (A 400nm 0.5 sup) in PBS, pH 7.4. Right panel: Antibody binding curves against PFs for soluble (A 400nm 0.5 sup) and insoluble (A 400nm 0.5 pellet) IgG conformers of heat-treated Avastin monomers, and for untreated Avastin and IVIg.

    Article Snippet: The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

    Techniques: Isolation, Size-exclusion Chromatography, Binding Assay

    IgG aggregates are primarily responsible for the enhanced anti-amyloid activities of Aβ- and Cibacron blue-isolated pAb IgGs. ( A ) Left panel: SEC chromatograms for ~0.5 mg/mL of Aβ-isolated IVIg IgGs, and for IVIg, untreated, or diluted into column elution buffer (0.1 M glycine, pH 2.7) that was used to elute Aβ-bound IVIg IgGs. SEC was carried out using a Superdex 200 increase 10/300 GL column (GE Healthcare) that was equilibrated with PBS, pH 7.4. Right panel: IgG binding curves against plate-immobilized PFs for untreated IVIg, and for Aβ column-isolated IVIg IgGs that were used unfractionated (Unfrac) or as SEC-isolated monomers (SEC Mon) or aggregates (SEC Aggs). SEC Aggs consisted of a pool of IgG conformers (dimers and HMW species) that eluted before the monomeric antibody. ( B ) Left panel: SEC chromatograms for ~0.5 mg/mL dye-isolated IVIg IgGs, and for unfractionated IVIg that was untreated or diluted into column elution buffer (PBS containing 1.5 M NaCl, pH 7.4) that was used to elute dye-bound IVIg IgGs. Right panel: IgG binding curves against plate-immobilized PFs for unfractionated and SEC-isolated conformers of dye-isolated IVIg IgGs, and for untreated IVIg.

    Journal: PLoS ONE

    Article Title: IgG Conformer's Binding to Amyloidogenic Aggregates

    doi: 10.1371/journal.pone.0137344

    Figure Lengend Snippet: IgG aggregates are primarily responsible for the enhanced anti-amyloid activities of Aβ- and Cibacron blue-isolated pAb IgGs. ( A ) Left panel: SEC chromatograms for ~0.5 mg/mL of Aβ-isolated IVIg IgGs, and for IVIg, untreated, or diluted into column elution buffer (0.1 M glycine, pH 2.7) that was used to elute Aβ-bound IVIg IgGs. SEC was carried out using a Superdex 200 increase 10/300 GL column (GE Healthcare) that was equilibrated with PBS, pH 7.4. Right panel: IgG binding curves against plate-immobilized PFs for untreated IVIg, and for Aβ column-isolated IVIg IgGs that were used unfractionated (Unfrac) or as SEC-isolated monomers (SEC Mon) or aggregates (SEC Aggs). SEC Aggs consisted of a pool of IgG conformers (dimers and HMW species) that eluted before the monomeric antibody. ( B ) Left panel: SEC chromatograms for ~0.5 mg/mL dye-isolated IVIg IgGs, and for unfractionated IVIg that was untreated or diluted into column elution buffer (PBS containing 1.5 M NaCl, pH 7.4) that was used to elute dye-bound IVIg IgGs. Right panel: IgG binding curves against plate-immobilized PFs for unfractionated and SEC-isolated conformers of dye-isolated IVIg IgGs, and for untreated IVIg.

    Article Snippet: The detection system consisted of an in house pan-Aβ reactive polyclonal rabbit antibody, AW7 [ ], a horse radish peroxidase-conjugated goat anti-rabbit IgG (GE healthcare), and TMB substrate (SureBlue ReserveTM; KPL).

    Techniques: Isolation, Size-exclusion Chromatography, Binding Assay

    Expression and characterization of TAP-tagged BRC motif in T. brucei (A) Map of the pLEW79FLR1TAP construct used for the in vivo doxycycline-inducible expression of a TAP-tagged single BRC motif (44 amino acids) with a 42 amino acid N-terminal flank. The recombinant protein (∼30 kDa) also has domains from calmodulin binding peptide (CBP, 26 amino acids), recognition site for TEV protease (TEV, 9 amino acids) and IgG-binding domains of Protein A (ProtA, 116 amino acids). Evaluation of the in vivo expression of the BRC motif-containing aptamer mRNA by RT-PCR (B) and corresponding protein by Western blot analysis (C). Since the wild-type procyclic T brucei cells (29-13) have tetracycline repressor protein (TetRep), the expression of the aptamer protein was inducible with doxycycline (Dox). RT-PCR products were generated with primers specific to TbBRCA2 BRC repeat. β-actin was used as a loading control in this analysis. Western Blot analysis was performed using a HRP-conjugated rabbit anti-goat IgG antibody. α-tubulin and the tet repressor were used as loading controls in this analysis. (D) Effect of expression of the BRC aptamer on the growth rate of procyclic forms of T. brucei . The controls included wild-type cells (WT 29-13) alone and wild-type cells transfected with the empty vector (Vector only). The other control is the recombinant cells grown in medium without doxycycline (No Dox cont). Results are mean ± SEM (n=12).

    Journal: Microbial pathogenesis

    Article Title: Peptide aptamer mimicking RAD51-binding domain of BRCA2 inhibits DNA damage repair and survival in Trypanosoma brucei

    doi: 10.1016/j.micpath.2010.11.007

    Figure Lengend Snippet: Expression and characterization of TAP-tagged BRC motif in T. brucei (A) Map of the pLEW79FLR1TAP construct used for the in vivo doxycycline-inducible expression of a TAP-tagged single BRC motif (44 amino acids) with a 42 amino acid N-terminal flank. The recombinant protein (∼30 kDa) also has domains from calmodulin binding peptide (CBP, 26 amino acids), recognition site for TEV protease (TEV, 9 amino acids) and IgG-binding domains of Protein A (ProtA, 116 amino acids). Evaluation of the in vivo expression of the BRC motif-containing aptamer mRNA by RT-PCR (B) and corresponding protein by Western blot analysis (C). Since the wild-type procyclic T brucei cells (29-13) have tetracycline repressor protein (TetRep), the expression of the aptamer protein was inducible with doxycycline (Dox). RT-PCR products were generated with primers specific to TbBRCA2 BRC repeat. β-actin was used as a loading control in this analysis. Western Blot analysis was performed using a HRP-conjugated rabbit anti-goat IgG antibody. α-tubulin and the tet repressor were used as loading controls in this analysis. (D) Effect of expression of the BRC aptamer on the growth rate of procyclic forms of T. brucei . The controls included wild-type cells (WT 29-13) alone and wild-type cells transfected with the empty vector (Vector only). The other control is the recombinant cells grown in medium without doxycycline (No Dox cont). Results are mean ± SEM (n=12).

    Article Snippet: Cell lysates containing equal amounts of protein were resolved by 4–12% SDS–PAGE, transferred to PVDF membrane and probed with a secondary rabbit IgG (anti-goat) antibody conjugated to HRP (1:1000) and detected by enhanced chemiluminescent technology (GE Healthcare, Brentwood, TN).

    Techniques: Expressing, Construct, In Vivo, Recombinant, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Generated, Transfection, Plasmid Preparation

    Localization of SERPINE2 in the human uterus . Longitudinal sections of the early secretory phase uterus (n = 5) on the slide were incubated with anti-mouse SERPINE2 antiserum and then treated with biotin-conjugated goat-anti-rabbit IgG and HRP-conjugated streptavidin (brown). For contrast, specimens were further stained with hematoxylin (blue). The representative picture is shown. Magnified pictures of the luminal epithelium (A), glandular epithelium (B), and myometrium (C) are shown. Bar = 50 μm. bv, blood vessel; ge, glandular epithelium; le, luminal epithelium; m, muscle; s, stroma.

    Journal: Reproductive Biology and Endocrinology : RB & E

    Article Title: SERPINE2, an inhibitor of plasminogen activators, is highly expressed in the human endometrium during the secretory phase

    doi: 10.1186/1477-7827-9-38

    Figure Lengend Snippet: Localization of SERPINE2 in the human uterus . Longitudinal sections of the early secretory phase uterus (n = 5) on the slide were incubated with anti-mouse SERPINE2 antiserum and then treated with biotin-conjugated goat-anti-rabbit IgG and HRP-conjugated streptavidin (brown). For contrast, specimens were further stained with hematoxylin (blue). The representative picture is shown. Magnified pictures of the luminal epithelium (A), glandular epithelium (B), and myometrium (C) are shown. Bar = 50 μm. bv, blood vessel; ge, glandular epithelium; le, luminal epithelium; m, muscle; s, stroma.

    Article Snippet: After gentle agitation in four changes of PBS for 15 min each, membranes were immunoreacted with horseradish peroxidase (HRP)-conjugated goat anti-rabbit immunoglobulin G (IgG) (1: 5000, GE Healthcare Life Sciences, Piscataway, NJ, USA), donkey anti-goat IgG (1: 3000, Jackson ImmunoResearch, West Grove, PA, USA), or horse anti-mouse IgG (1:3000, Cell Signaling Technology, Beverly, MA, USA), respectively, in blocking solution for 2 h at 37°C.

    Techniques: Incubation, Staining