Clicking the images or links will redirect you to a website hosted by BenchSci that provides third-party scientific content. Neither the content nor the BenchSci technology and processes for selection have been evaluated by us; we are providing them as-is and without warranty of any kind, including for use or application of the Thermo Fisher Scientific products presented.    Western blot analysis was performed on whole cell extracts (30 µg lysate) of K-562 (Lane 1) and U87-MG (Lane 2). The blots were probed with Anti-SOD2 Mouse Monoclonal Antibody (Product # MA1-106, 0.25 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 31430) at dilutions 1:5,000 (Fig. 1), 1:100,000 (Fig. 2) and 1:200,000 (Fig. 3). A 25 kDa band corresponding to SOD2 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody after blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).   Western blot analysis of beta-3 Tubulin was performed by loading 25 µg of various brain, SHSY5Y, and HeLa cell lysates and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a beta-3 Tubulin monoclonal antibody (Product # MA1-118) at a dilution of 1:2000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Note: analysis indicates both reactive (neuronal) and non-reactive negative control (HeLa cell lysate) for beta-3 Tubulin using Product # MA1-118.   Western blot analysis of GSK-3 was performed by loading 20 µg of the indicated whole cell lysates and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BOX ). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. GSK-3 (alpha and beta isoforms) was detected at ~51 kD and ~47 kD using a GSK-3 monoclonal antibody (Product # MA3-038) at a dilution of 1:1000 in 5% Milk in TBST overnight at 4C on a rocking platform, followed by a Goat anti-Mouse IgG Secondary Antibody, HRP conjugate (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal Pico substrate (Product # 34078) and the myECL Imager (Product # 62236).   Western blot analysis of CD38 was performed by loading 50 µg of the indicated cell lysates prepared using IP Lysis Buffer (Product # 87788) or Mem-PER Plus Membrane Protein Extraction Kit (Product # 89842), and 10 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BOX ). Proteins were transferred to a nitrocellulose membrane (Product # 88014) using the Power Blotter (Product # 22834), and blocked with 5% milk in TBST for at least 1 hour at room temperature. CD38 was detected at ~45 kD using a CD38 monoclonal antibody (Product # MA1-182) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:40,000 for at least 30 minutes. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).   Western blot analysis of MEK1 was performed by loading 20 µg of the indicated whole cell lysates per well and 5 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% milk in TBST for 1 hour at room temperature. MEK1 was detected at ~43 kD using a MEK1 monoclonal antibody (Product # 13-3500) at a dilution of 1 µg/mL in 5% milk in TBST overnight at 4C on a rocking platform. After washing, blots were probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico substrate (Product # 34080).   Western blot analysis of eIF2S1 was performed by loading 80 µg of the indicated cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an eIF2S1 monoclonal antibody (Product # MA1-079) at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).   Immunohistochemistry was performed on paraffin-embedded human colon cancer tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA (Product # 37525) in PBST for 30 minutes at room temperature and then probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:100 for 1 hour in a humidified chamber. Tissues were washed extensively with PBS/0.025% Tween-20 (Product # 28314) and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000) for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:500 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axiovision microscope at 40X magnification.   Immunohistochemistry was performed on deparaffinized canine fetal brain tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95°C. Following antigen retrieval tissues were blocked with Avidin/Biotin Blocking kit (Product # 00-4303) at room temperature and then probed with a Sox2 monoclonal antibody (Product # MA1-014) at a dilution of 1:200 for one hour at room temperature. Tissues were washed extensively with TBS + 0.025% Triton X-100 (Product # 28314). Detection was performed using a goat anti-mouse HRP secondary antibody (Product # 31430) at a dilution of 1:500 followed by colorimetric detection using metal enhanced DAB (Product # 34065). Tissues were counterstained with hematoxylin and prepped for mounting. Images were taken on a Zeiss Axiovision microscope at 20X magnification. The bottom layer with a dense staining of Sox2 is in the subependymal plate, where there are large numbers of progenitor cells. The upper layer is neuroparenchyma where cells are differentiated, therefore less staining of Sox2. Note: Data courtesy of Innovators Program.   Immunohistochemistry was performed on paraffin-embedded human breast cancer tissue sections. To expose target proteins, heat-induced epitope retrieval was performed using 10mM sodium citrate (pH 6.0) buffer for 20 minutes at 95ºC. Following antigen retrieval, tissues were blocked in 3% BSA (Product # 37525) in PBST for 30 minutes at room temperature and then probed with an Ezrin monoclonal antibody (Product # MA5-13862) at a dilution of 1:100 for 1 hour in a humidified chamber. Tissues were washed extensively with PBS/0.025% Tween-20 (Product # 28314) and endogenous peroxidase activity quenched with Peroxidase Suppressor (Product # 35000) for 30 minutes at room temperature. Detection was performed using an HRP-conjugated goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:500 followed by colorimetric detection using Metal Enhanced DAB Substrate Kit (Product # 34065). Images were taken on a Zeiss Axiovision microscope at 40X magnification.   Indirect ELISA analysis of Digoxin was performed by preparing a 10 µg/mL solution of BSA (Product # 23209) or Digoxin-labeled BSA in 0.2M Carbonate-Bicarbonate, pH 9.4 (Product # 28382). 100uL of each solution was added to separate wells of a clear 96-well plate (Product # 15041), and incubated overnight at 4C. Each well was blocked with SuperBlock in PBS (Product # 37515). The plate was then washed with PBST and incubated with 100uL per well of Digoxin monoclonal antibody (Product # MA5-14745) in triplicate at ~1000 - 2pg/mL for 1 hour at room temperature. The plate was washed with PBST and incubated with 100 µL per well of goat anti-mouse IgG-HRP secondary antibody (Product # 31430) in all test wells at 1:25,000 for 30 minutes at room temperature. Detection was performed using 1-Step TMB Ultra substrate (Product # 34028) for 15 minutes at room temperature. The plate was then stopped with 0.2M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.   Direct ELISA analysis of N-terminal and C-terminal His-tagged proteins was performed by coating wells of a plate with a recombinant N-terminal His-tagged protein (left panel) or a recombinant C-terminal His-tagged protein (right panel) at a concentration of 5 µg/mL overnight at 4C. The plate was washed 3 times with ELISA Wash Buffer (Product # N503), 100 µL of a 6x-His Epitope Tag monoclonal antibody (Product # MA1-135) was added to wells in duplicate at 1250, 625, 312.5, 156, 78, 39, 19.5, 9.5, 4.5 and 0 ng/mL concentrations, and the samples were incubated for 2 hours at room temperature. The plate was washed, and then incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG secondary antibody (Product # 31430) at a dilution of 1:10,000 for 1 hour at room temperature, and washed again with ELISA Wash Buffer. The plate was developed by incubating 100 µL per well of 1-Step Ultra TMB substrate (Product # 34028) per well for 10 minutes at room temperature in the dark. The reaction was stopped with 100 µL per well of 0.16M sulfuric acid. Absorbances were read on a spectrophotometer at 450-550 nm.   Immunoprecipitation of HuR was performed using MCF-7 cells. Antigen-antibody complexes were formed by incubating 500 µg of MCF-7 whole cell lysates with 2 µg of HuR (3A2) monoclonal antibody (Product # MA1-167) overnight on a rocking platform at 4C. The immune complexes were captured on 100 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). The IP eluate (right lane) and 20 µg of MCF-7 lysate (left lane) were resolved on a 4-20% Tris-Glycine polyacrylamide gel, transferred to a nitrocellulose membrane, and blocked with 5% Milk in TBST for at least 1 hour. The membrane was probed with a HuR (3A2) monoclonal antibody monoclonal antibody (Product # MA1-167) at a dilution of 1:250 overnight rotating at 4C, washed in TBST, and probed with goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 30 minutes at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).   Immunoprecipitation of PKC alpha was performed using Jurkat whole cell lysate. Antigen-antibody complexes were formed by incubating 400 µg of lysate with 5 µg of a PKC alpha monoclonal antibody (Product # MA1-157) overnight on a rocking platform at 4°C. The immune complexes were captured on 50 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Jurkat cell lysate (40 µg) was loaded as a positive control (left lane). The sample was resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for 1 hour. The membrane was probed with a PKC alpha monoclonal antibody (Product # MA1-157) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). Product # 31430 has been successfully used in Western blot, IHC and IP applications.rrProduct # 31430 reacts with the heavy chains of mouse igg and with the light chains common to most mouse immunoglobulins, but does not react against non-immunoglobulin serum proteins. However, this antibody may cross-react with immunoglobulins from other species.rrStore product at 4°C until opened. To extend the shelf-life of this product, add an equal volume of glycerol to make a final concentration of approximately 50% glycerol and store at -20°C.rrReconstitute with 2.0 mL of distilled water (0.8 mg/mL after restoration). Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents. Host server : magellan-srch-1-prod-green:8080/10.253.224.248:8080. git-commit: 2cd8645d2fc6bfe4ccb4abfa14772b0a94f68e98 git-url: http://victoria.invitrogen.com:8333/magellan/core.git git-branch: origin/release/1.27.0-2021.08.32-1.0