Cells grown on coverslips were fixed with 2% paraformaldehyde and permeabilized with 0.5% NP-40. Immunofluorescence staining was carried out by incubating with an anti-BLM antibody (Santa Cruz Biotechnology, sc-7790, 1:150), an anti-RPA34 antibody (GeneTex, clone 9H8, 1:500), an anti-PICH antibody (Abnova, H54821-B01P, 1:500), or an anti-TRF1 antibody (GeneTex, clone 4E4, 1:500), followed by secondary antibody conjugated with respective Alexa Fluorophores (Molecular Probes, 1:500). The cells were fixed again with 4% paraformaldehyde and dehydrated by successive incubation in 70, 95 and 100% ethanol before subjected to FISH analysis. PNA probes for FISH analysis were TMR-, Cy5- or Alexa488-OO-5′-(CCCTAA)3-3′ (telomeric sequence). DNA was stained by 0.1 μg ml−1 DAPI. Coverslips were mounted onto glass slides in Prolong Gold Antifade Reagent (Invitrogen). Images were acquired with a Nikon Ti-U microscope using a × 100 objective and collected as a stack of 0.2-μm increments in the z axis. Image deconvolution was conducted using the AutoQuant X3 software. Unless otherwise noted, images were viewed as a single section on the z axis.
Anti blm antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-BLM antibody is a reagent used in laboratory research. It is designed to detect and bind to the BLM protein, which plays a role in DNA repair processes. This antibody can be used in various experimental techniques, such as Western blotting, immunoprecipitation, and immunofluorescence, to study the expression and localization of the BLM protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ti u microscope, by Nikon (1 mentions)
Alexa fluorophore, by Thermo Fisher Scientific (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene difluoride membrane, by GE Healthcare (1 mentions)
Irdye 800cw donkey anti goat igg secondary antibody, by LI COR (1 mentions)
Primary monoclonal antibody against β actin, by Proteintech (1 mentions)
2 protocols using anti blm antibody
1
Immunofluorescence and FISH Protocol
2
Western Blot Analysis of BLM Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein was extracted from the cells or tissues using lysis buffer containing protease inhibitors. Proteins (30 µg) from each sample were loaded on 10% SDS-polyacrylamide gels for electrophoresis. Following electrophoresis, the proteins were transferred onto polyvinylidene difluoride membranes (GE Healthcare, Chalfont, UK), which were then blocked with 10% non-fat milk in Tween/Tris-buffered salt solution (TTBS; 20 mm Tris-Cl, pH 7.5, 0.15 M NaCl and 0.05% Tween-20) for 1 h. Following incubation with the anti-BLM antibody (Santa Cruz Biotechnology, Inc., Dallas, TX, USA) at 4°C overnight, the membrane was washed and incubated with IRDye® 800CW donkey anti-goat IgG secondary antibody at room temperature for 2 h followed by LI-COR Odyssey gel imaging scanner detection (LI-COR Biosciences, Lincoln, NE, USA). To verify equal loading of protein, the blots were reprobed with primary monoclonal antibody against β-actin (ProteinTech Group, Inc., Chicago, IL, USA).
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