H 175

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The H-175 is a laboratory centrifuge designed for high-speed separation of biological samples. It features a maximum speed of 17,500 rpm and can accommodate a variety of sample volumes and rotor configurations. The H-175 is intended for general laboratory use in the life sciences and biotechnology industries.

Automatically generated - may contain errors

Lab products found in correlation

Ecl detection kit, by Cytiva (2 mentions) Dynabeads protein a, by Thermo Fisher Scientific (1 mentions) Mab1920, by Merck Group (1 mentions) Axioskop 2 microscope, by Zeiss (1 mentions) Alexa 488 conjugated goat anti rat, by Thermo Fisher Scientific (1 mentions) Alexa 594 conjugated goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Las 1000 system, by Fujifilm (1 mentions) Bullet blender, by Next Advance (1 mentions) Protease inhibitors cocktail, by Roche (1 mentions)

9 protocols using h 175

Immunoblot Analysis of Kv11.1a and Kv11.1a-USO Proteins

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblot analysis was performed as previously described [10 (link)]. The cell lysates were subjected to SDS-polyacrylamide gel electrophoresis and then electrophoretically transferred onto nitrocellulose membranes. The membranes were incubated with an anti-Kv11.1 antibody against the N-terminus of Kv11.1a and Kv11.1a-USO proteins (H-175, Santa Cruz, Santa Cruz, CA) at a 1:600 dilution and visualized with the ECL detection kit (Amersham, Piscataway, NJ). The expression level of hygromycin B phosphotransferase (HPH) encoded by hygromycin B resistance gene was used as loading control [6 (link)].

Gong Q., Stump M.R, & Zhou Z. (2014). Upregulation of Functional Kv11.1 Isoform Expression by Inhibition of Intronic Polyadenylation with Antisense Morpholino Oligonucleotides. Journal of molecular and cellular cardiology, 0, 26-32.

+ Open protocol

+ Expand

RNase Protection Assay and Immunoblot for hERG Expression

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNase protection assay (RPA) was performed as previously described (Gong et al., 2011 (link)). Briefly, antisense RNA riboprobes were transcribed in vitro in the presence of biotin-14-CTP. The probe contained 277 nt spanning the region of exons 12 and 13. The total length of the probe was 409 nt and contained sequences from the pCRII vector at both ends. The expression level of hygromycin B resistance gene in the pcDNA5 vector was used as an internal control for normalization. The probe for the hygromycin B resistance gene contained 158 nt of the gene and 70 nt from the pGEM vector. Yeast RNA was used as a control for the complete digestion of the probes by RNase. The relative intensity of RPA bands was quantified using ImageJ.
Immunoblot was performed as previously described (Gong et al., 2010 (link)). The cell lysates were subjected to SDS-polyacrylamide gel electrophoresis and the blots were probed with an antibody against the N-terminus of the hERG channel protein (H175, Santa Cruz Biotechnology Inc., Santa Cruz, CA). The expression level of hygromycin B phosphotransferase (HPH) encoded by hygromycin B resistance gene in the pcDNA5 vector was used as an internal control for normalization. The polyclonal antibody against HPH was used at 1:1000 dilution as previously described (Gong et al., 2010 (link)).

Gong Q., Stump M.R, & Zhou Z. (2014). Position of Premature Termination Codons Determines Susceptibility of hERG mutations to Nonsense-Mediated mRNA Decay in Long QT Syndrome. Gene, 539(2), 190-197.

+ Open protocol

+ Expand

Immunoprecipitation of NEO1 and LMγ1

Check if the same lab product or an alternative is used in the 5 most similar protocols

SK-N-SH cells were used to prepare cell extracts with a buffer containing 20 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, and protease inhibitors by 5 min incubation on ice. Samples were centrifuged at 13,000 × g by 1 min at 4°C, and supernatants (1000 μg total protein) were immunoprecipitated with Dynabeads protein A (Thermofisher) bead-immobilized antibodies for 1h. NEO1 was immunoprecipitated with 2 μg of a rabbit polyclonal antibody (H-175, Santa Cruz) and LMγ1 was immunoprecipitated with 2 μg of mouse monoclonal antibody (MAB1920, Millipore). Immunoprecipitated samples were solubilized in loading buffer with ß-mercaptoethanol, and analyzed by Western blot as indicated in (Figure 3).

Villanueva A.A., Puvogel S., Lois P., Muñoz-Palma E., Ramírez Orellana M., Lubieniecki F., Casco Claro F., Gallegos I., García-Castro J., Sanchez-Gomez P., Torres V.A, & Palma V. (2018). The Netrin-4/Laminin γ1/Neogenin-1 complex mediates migration in SK-N-SH neuroblastoma cells. Cell Adhesion & Migration, 13(1), 33-40.

+ Open protocol

+ Expand

Immunoblot Analysis of Kv11.1a and Kv11.1a-USO Isoforms

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblot analysis was performed as previously described [7 (link)]. Kv11.1a and Kv11.1a-USO proteins were detected using an anti-Kv11.1 antibody directed against the N-terminus of the isoforms (H-175, Santa Cruz, CA, USA). The relative expression levels of Kv11.1a and Kv11.1a-USO were normalized to the expression of hygromycin B phosphotransferase (HPT) encoded by the hygromycin B resistance gene [7 (link)]. PABPN1 was detected using an anti-PABPN1 antibody (Abcam, Cambridge, MA, USA). The expression of β-tubulin was used as a loading control as previously described [10 (link)]. The intensity of each protein band was quantified using Image J software.

Stump M.R., Nguyen R.T., Drgastin R.H., Search D., Gong Q, & Zhou Z. (2021). Regulation of Kv11.1 Isoform Expression by Polyadenylate Binding Protein Nuclear 1. International Journal of Molecular Sciences, 22(2), 863.

+ Open protocol

+ Expand

Quantification of Kv11.1 Channel Surface Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

The surface expression of Kv11.1 channels containing an external HA epitope tag was determined by immunofluorescence as described by Ficker et al.^{17 (link)} Briefly, live cells were blocked with PBS containing 5% goat serum, probed with rat monoclonal anti-HA antibody for 1 hour at 4°C (3F10, Roche, 1:100). Cells were washed with PBS and fixed with 4% paraformaldehyde at 4°C. Fixed cells were re-blocked with PBS containing 5% goat serum and permeabilized with 0.1% Triton X-100. Permeabilized cells were re-probed with rabbit anti-Kv11.1 N-terminus antibody at 22–23°C (H175, Santa Cruz Biotechnology Inc., 1:300). Cells were washed and incubated with Alexa 488-conjugated goat anti-rat and Alexa 594-conjugated goat anti-rabbit antibody (Molecular Probes, Eugene, OR). Images were acquired with a Zeiss Axioskop 2 microscope.

Gong Q., Stump M.R., Deng V., Zhang L, & Zhou Z. (2014). Identification of Kv11.1 Isoform Switch as a Novel Pathogenic Mechanism of Long QT Syndrome. Circulation. Cardiovascular genetics, 7(4), 482-490.

+ Open protocol

+ Expand

Immunoblot Analysis of Kv11.1a and Kv11.1a-USO Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunoblot analysis was performed as previously described (Gong et al., 2010 (link)). The cell lysates were subjected to SDS-polyacrylamide gel electrophoresis and then electrophoretically transferred onto nitrocellulose membranes. The membranes were incubated with an anti-Kv11.1 antibody against the N-terminus of Kv11.1a and Kv11.1a-USO proteins (H-175, Santa Cruz, Santa Cruz, CA) at a 1:600 dilution and visualized with the ECL detection kit (Amersham, Piscataway, NJ). The expression level of hygromycin B phosphotransferase (HPT) encoded by the hygromycin B resistant gene was used as loading control (Gong et al., 2010 (link)).

Gong Q., Stump M.R, & Zhou Z. (2017). Upregulation of functional Kv11.1a isoform expression by modified U1 small nuclear RNA. Gene, 641, 220-225.

+ Open protocol

+ Expand

Immunoblotting for CYP2W1 detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human and mice tissues were homogenized using the Bullet Blender (Next Advance, Averill Park, NY, USA) in a buffer containing 10 mM Tris–HCl pH 7.5, 0.5 mM EDTA, 0.25 M sucrose with addition of protease inhibitors cocktail (Roche Diagnostics, Mannheim, Germany). HCC2998 cells were lysed in RIPA buffer containing protease inhibitors cocktail for 30 min at 4°C. Denatured protein (40 μg) samples were run on 10% SDS-PAGE, transferred to nitrocellulose membranes and immunoblotted with two different rabbit anti-human CYP2W1 antibodies (H175, Santa Cruz, CA, USA, dilution 1:200 and the C-terminal in-house anti-CYP2W1-852 [4 (link)], dilution 1:1000), rabbit anti-mouse CYP2W1 antibodies (in-house anti-CYP2W1-3675, dilution 1:500), or with rabbit anti-ERp29 (in-house antibody reacting with both species [15 (link)], dilution 1:1000),javascript:void(0); followed by goat anti-rabbit conjugated horseradish peroxidase secondary antibodies (Dako, Glostrup, Denmark, dilution 1:2000). Filters were developed using SuperSignal West Femto Chemiluminescent Substrate (Pierce, Rockford, IL, USA) and signals detected by LAS-1000 system (Fujifilm, Japan).

Choong E., Guo J., Persson A., Virding S., Johansson I., Mkrtchian S, & Ingelman-Sundberg M. (2015). Developmental Regulation and Induction of Cytochrome P450 2W1, an Enzyme Expressed in Colon Tumors. PLoS ONE, 10(4), e0122820.

+ Open protocol

+ Expand

Western Blot Analysis of Kv11.1 Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were washed with PBS and lysed with lysis buffer (50 mmol/L Tris-HCl, pH 7.4 containing 150 mmol/L NaCl, 1% Triton X-100) plus a protease inhibitor cocktail (100 µmol/L phenylmethylsulfonyl fluoride, 1 µg/ml pepstatin A, 1 µg/ml of leupeptin, 2 µl/ml of aprotinin). The cell lysates were subjected to SDS-polyacrylamide gel electrophoresis and then electrophoretically transferred onto nitrocellulose membranes. After transfer, the membranes were blocked with 5% nonfat dry milk and 0.2% Tween 20 in PBS for 1 h. The membranes were probed with a 1:600 dilution of an anti-Kv11.1 antibody directed against the N-terminus of the protein (H175, Santa Cruz Biotechnology Inc., Santa Cruz, CA) or a 1:1000 dilution of an anti-HA antibody (HA.11, Convance, Berkeley, CA), and visualized with horseradish peroxidase-conjugated second antibody and ECL detection kit. The expression level of hygromycin B phosphotransferase encoded by the hygromycin B resistance gene was used as a loading control. The polyclonal antibody against hygromycin B phosphotransferase was custom-generated by Genscript (Piscataway, NJ) and used at a 1:1000 dilution.^{11 (link)} The intensity of the protein bands was quantified using ImageJ.

+ Open protocol

+ Expand

Netrin-4 Binding Partners Identification

Check if the same lab product or an alternative is used in the 5 most similar protocols

SK-N-SH cells were incubated with human recombinant Netrin-4 (200 ng/ml) for 1h. Later, cell extracts were prepared in a buffer containing 20 mM Tris, pH 7.4, 150 mM NaCl, 1% NP-40, and protease inhibitors by 5 min incubation on ice. Samples were centrifuged at 13,000 × g by 1 min at 4°C, and supernatants (1000 μg total protein) were immunoprecipitated with protein A/G bead-immobilized antibodies for 1h. NEO1 was immunoprecipitated with 5 μg of a rabbit polyclonal antibody (H-175 Santa Cruz) and NTN4 was immunoprecipitated with 5 μg of NTN4 goat polyclonal antibody. Immunoprecipitated samples were solubilized in loading buffer with ß-mercaptoethanol, and analyzed by Western blot as indicated in the Figure 1.

Villanueva A.A., Falcón P., Espinoza N., Luis S.R., Milla L.A., Hernandez-SanMiguel E., Torres V.A., Sanchez-Gomez P, & Palma V. (2016). The Netrin-4/ Neogenin-1 axis promotes neuroblastoma cell survival and migration. Oncotarget, 8(6), 9767-9782.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!