Carbo free blocking solution sp 5040

Manufactured by Vector Laboratories

Sourced in United States

Carbo-Free Blocking Solution (SP-5040) is a protein-free blocking solution designed for use in immunoassays and Western blotting applications. It is formulated to reduce non-specific background signals without interfering with the target analyte.

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Lab products found in correlation

Anti piezo1, by Thermo Fisher Scientific (1 mentions) Lsm 880 airyscan, by Zeiss (1 mentions) Vectashield hardset mounting media, by Vector Laboratories (1 mentions) Ulex europaeus agglutinin, by Vector Laboratories (1 mentions) Pha l, by Vector Laboratories (1 mentions) Annexin 5, by BioLegend (1 mentions) Streptavidin pe, by BD (1 mentions)

Spelling variants of this product

Carbo-Free Blocking Solution (66 citations) Blocking solution (11 citations) SP-5040 (6 citations)

3 protocols using carbo free blocking solution sp 5040

Immunostaining of Piezo1 in Aortic Smooth Muscle Cells

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Freshly isolated mouse aortic smooth muscle cells and HAoSMCs were seeded onto glass coverslips and kept at 37°C for 1–2 h while attached to the surface of the coverslips. After fixed in 4% PFA and repeated washing in glicine-PBS solution (100 mM, 15 min, 22°C), cells were permeabilized with Triton-X-100 (0.5 v/v%, 10 min, 22°C). After PBS washing (3x) non-specific biding sites were blocked with Carbo-Free Blocking Solution (SP-5040, VECTOR Laboratories, Burlingame, CA, United States). Immunostaining was performed using anti-Piezo1 (Thermo Fisher Scientific, Rockford, IL, United States, MA5-32876, mouse-IgG, monoclonal) primary antibody with Cy3 anti-mouse secondary antibody labelling (A10521, Life technologies, Eugene, OR, United States). Images were taken using Zeiss laser scanning confocal microscope (Zeiss LSM880 Airyscan; Zeiss, Oberkochen, Germany) using 405, 488 and 543 nm excitation wavelengths and 10x or 63x oil immersion objective. On HAoSMCs anti-Piezo1 and anti-alfa smooth muscle specific actin (αSMA) (Thermo Fisher Scientific, Rockford, IL, United States, MA5-32876, mouse-IgG, monoclonal; PA516697, rabbit-IgG, polyclonal, respectively) primary antibodies were applied.
Paraffin embedded human aorta samples were subjected to deparaffinization process, and the protocol described above was applied as staining procedure.

Szabó L., Balogh N., Tóth A., Angyal Á., Gönczi M., Csiki D.M., Tóth C., Balatoni I., Jeney V., Csernoch L, & Dienes B. (2022). The mechanosensitive Piezo1 channels contribute to the arterial medial calcification. Frontiers in Physiology, 13, 1037230.

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Lectin Staining of Liver Tissue

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Liver sections (5 μm thick) from control and LDKO mice were cut, deparaffinized, and stained for lectins such as UEA (Ulex Europaeus Agglutinin) (binds fucose), PHA-L (binds complex glycans), and SNA (binds sialic acid) from Vector Laboratories. Briefly, sections were blocked with Carbo-free Blocking solution (SP-5040, Vector Laboratories) for 30 min at room temperature, followed by fluorescently labeled lectin in PBS for 30 min at room temperature. The slides were then washed twice in PBST (Phosphate Buffered Saline with Triton X-100) and stained, mounted with Vectashield Hardset mounting media (H-1400, Vector Laboratories), and imaged using confocal microscope LSM 710 at Institute for Genomic Biology Core facility, UIUC using 488-nm laser, and a red psuedocolor was assigned to it.

Mathur B., Shajahan A., Arif W., Chen Q., Hand N.J., Abramowitz L.K., Schoonjans K., Rader D.J., Kalsotra A., Hanover J.A., Azadi P, & Anakk S. (2021). Nuclear receptors FXR and SHP regulate protein N-glycan modifications in the liver. Science Advances, 7(17), eabf4865.

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Whole-Blood Flow Cytometry for RBCs

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Whole‐blood flow cytometry assays were used for Figures 1A, 2A–C, 2E and 3A–C. Purified RBCs were used for Figures 2D and 4C,D. RBC gating was applied by forward and side scatter gating of both whole blood and purified RBC flow cytometry experiments.¹⁷ Staining with anti‐glycophorin A (GPA) confirmed gates contained >99% RBCs (data not shown). For lectin flow cytometry, ~5 × 10⁶ RBCs were washed three times in phosphate‐buffered saline and incubated for 10 min at room temperature in calcium buffer (10 mM HEPES, 150 mM NaCl₂, 2.5 mM CaCl₂, pH 7.4) containing 10% Carbo‐Free Blocking Solution (SP5040; Vector Laboratories), then stained in the dark for 30 min. Annexin V staining was carried out according to the manufacturer's instructions (640945; BioLegend). For biotinylated lectin staining, cells were then washed and incubated with streptavidin PE (0.67 μg/ml, 554 061; BD Pharmingen) for 30 min at room temperature in blocking buffer.

Cao H., Mathur A., Robertson C., Antonopoulos A., Henderson S., Girard L., Wong J.H., Davie A., Wright S., Brewin J., Rees D.C., Dell A., Haslam S.M, & Vickers M.A. (2022). Measurement of erythrocyte membrane mannoses to assess splenic function. British Journal of Haematology, 198(1), 155-164.

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