Protein block solution

Manufactured by Abcam

Sourced in United Kingdom

Protein block solution is a laboratory reagent used to prevent non-specific binding of proteins in immunoassays, such as Western blotting and ELISA. It contains a mixture of proteins and detergents that help to block unoccupied binding sites on the solid support, reducing background signal and improving the specificity of the assay.

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Lab products found in correlation

Axiovert 200m, by Zeiss (2 mentions) Microtome, by Leica (1 mentions) Primary antibody against cleaved caspase 3, by Cell Signaling Technology (1 mentions) Proteinase k, by Agilent Technologies (1 mentions) Envision kit, by Agilent Technologies (1 mentions) Cytation 5, by Agilent Technologies (1 mentions) Light microscopy, by Olympus (1 mentions) Dab substrate, by Abcam (1 mentions) Cell dissociation reagent, by Thermo Fisher Scientific (1 mentions) Antibodies against vimentin, by Cell Signaling Technology (1 mentions) Ecl reagent kit, by GE Healthcare (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions) Mounting medium, by Abcam (1 mentions) Hematoxylin solution, by Abcam (1 mentions) Mouse and rabbit specific hrp dab abc detection ihc kit, by Abcam (1 mentions) Signalstain antibody diluent, by Cell Signaling Technology (1 mentions) Triton x 100, by Thermo Fisher Scientific (1 mentions)

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Protein Block

4 protocols using protein block solution

Histological Analysis of Tumor Sections

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The histopathological study was conducted with the hematoxylin and eosin (H&E) stain^{41 (link)}. Mice were subjected to euthanasia by transcardial perfusion with 4% paraformaldehyde in PBS and stored in the 70% ethanol solution. Tumors were blocked in longitudinal sections and processed for paraffin embedding, followed by cutting with a microtome (Leica Microsystems Inc., IL, USA). H&E stains were applied to tumor sections with the thickness of 5 μm, and the images were photographed using a light microscopy (Olympus).
The immunohistochemical (IHC) analysis of cleaved caspase-3 was carried out following the previous report^{44 (link)}. In brief, the retrieval of antigens was performed using Proteinase K (Dako, CA, USA). Samples were permeabilized with 0.05% Triton-X in PBS for 5 min. Protein Block Solution (Abcam, MA, USA) was applied for non-specific antigen blocking. Sample slides were then incubated with the primary antibody against cleaved caspase-3 (Cell Signaling Tech.), and the result was analyzed by using the EnVision kit (Dako). Slides were mounted in MM24 (Leica, Germany). Digital images of whole-tissue sections were acquired using a Cytation 5 (Biotek, VT, USA). Ten images from each slide were selected at random at 40 × magnification using GEN5 software (Biotek).

Chiang C.J, & Hong Y.H. (2021). In situ delivery of biobutyrate by probiotic Escherichia coli for cancer therapy. Scientific Reports, 11, 18172.

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Epithelial-Mesenchymal Transition Induction

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Fetal bovine serum (FBS), Cell Dissociation Reagent, medium LHC-9 and Dulbecco’s modified Eagle medium (DMEM) were provided by Invitrogen (Carlsbad, CA). The antibody against VECTOR ImmPRESS Anti-Goat Ig was provided by Santa Cruz Biotechnology (Santa Cruz, CA). Protein Block solution and DAB substrate were provided by Abcam (Cambridge, MA). Antibodies against vimentin, E-cadherin, N-cadherin and TCTP were purchased from Cell Signaling (Boston, MA). ECL reagent kit was purchased from GE Healthcare Life Sciences (Piscataway, NJ). NNK was purchased from Chemsyn Science Laboratories (Lenexa, KS). 10 µM NNK was used in the experiments [20 (link)]. PM_2.5 was collected at Kowloon Tong of Hong Kong and 5 µg/ml PM_2.5 was used in the experiments as details described in our previous study [21 (link)].

Liu L.Z., Wang M., Xin Q., Wang B., Chen G.G, & Li M.Y. (2020). The permissive role of TCTP in PM2.5/NNK-induced epithelial–mesenchymal transition in lung cells. Journal of Translational Medicine, 18, 66.

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Immunohistochemical Staining of Tissue Samples

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The specimen sections were deparaffinized and rehydrated. Then, endogenous peroxidase was blocked using a hydrogen peroxide block (Abcam, UK). The specimens were washed, and antigen retrieval was performed by heating in a 10 mM citrate buffer. After protein block solution (Abcam, UK) was applied to reduce nonspecific background staining, the specimens were incubated with primary antibodies overnight at room temperature in a humidified chamber. The antigen-antibody complex was then detected using a Mouse and Rabbit Specific HRP/DAB (ABC) Detection IHC Kit (Abcam, UK), following the manufacturer's instructions. The sections were counterstained with hematoxylin solution (Mayer's modified, Abcam, UK) and dehydrated before mounting with mounting medium (Abcam, UK). As a negative control, the primary antibody was replaced with SignalStain antibody diluent (Cell Signaling Technology, USA). The stained specimens were examined under a Motic BA210 microscope.

Leelawat S., Leelawat K., Yimsoo T., Wunnakup T., Monton C., Khamthong N., Madaka F., Maha A, & Songsak T. (2022). Antitumor Effects of Delta (9)-Tetrahydrocannabinol and Cannabinol on Cholangiocarcinoma Cells and Xenograft Mouse Models. Evidence-based Complementary and Alternative Medicine : eCAM, 2022, 6477132.

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Immunocytochemical Analysis of Muscle Progenitor Cells

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Cells were fixed in 4% paraformaldehyde and permeabilized at room temperature either in 1:1 methanol:acetone solution for 10 min (for MYHC) or in 0.5% Triton‐X‐100 (ThermoFisher Scientific) for 15 min (for PAX7 and MYOD). Subsequently, nonspecific antibody binding was blocked by incubating the samples at room temperature in either protein block solution (Abcam, Cambridge, UK) for 1 h or in homemade blocking solution (10% goat serum, 2% BSA, 0.25% Triton® X-100 in 1× PBS) for 30 min, respectively. Primary antibodies (Table 1) were then incubated for 16 h at 37 °C (PAX7) or 4 °C overnight (MYOD and MYHC) in blocking solution (MYOD and PAX7) or antibody diluent reagent (MYHC; Life Technologies). After washing in PBS or PBS plus 0.05% Triton X (PBST), MDPCs were incubated with secondary AF488-conjugated goat anti-mouse antibody for 1 h. Cells were washed in PBS or PBST for a further 3 times and overlayed with mountant containing DAPI (Sigma‐Aldrich) and subsequently covered with a coverslip. Images were captured on Axiovert 25 and Axiovert 200M inverted microscopes and analysed with Zen Software (Zeiss, Oberkochen, Germany). No antibody (unstained) and secondary antibody only controls were used (Supplementary Fig. 3).

Dan-Jumbo S.O., Riley S.E., Cortes-Araya Y., Ho W., Lee S., Thrower T., Esteves C.L, & Donadeu F.X. (2024). Derivation and long-term maintenance of porcine skeletal muscle progenitor cells. Scientific Reports, 14, 9370.

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