Quickblock primary antibody dilution buffer for western blot

Manufactured by Beyotime

Sourced in China

QuickBlock™ Primary Antibody Dilution Buffer is a ready-to-use solution designed for the dilution of primary antibodies used in Western blot analysis. It is formulated to maintain the stability and specificity of the primary antibody during the dilution process.

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Lab products found in correlation

Quickblock blocking buffer for western blot, by Beyotime (5 mentions) Quickblock secondary antibody dilution buffer for western blot, by Beyotime (3 mentions) Pvdf membrane, by Merck Group (2 mentions) Ipvh00010, by Merck Group (2 mentions) Chemidoc touch chemiluminescence imaging system, by Bio-Rad (1 mentions) Ab125066, by Abcam (1 mentions) Chemidoc xrs imaging system, by Bio-Rad (1 mentions) Ab46545, by Abcam (1 mentions) Anti β actin af5003, by Beyotime (1 mentions) Hrp labelled goat anti rabbit igg h l, by Beyotime (1 mentions) Protein extraction kit, by Solarbio (1 mentions) Gapdh rabbit mab, by ABclonal (1 mentions) Hrp labeled goat anti rabbit igg h l a0208, by Beyotime (1 mentions) Bradford protein assay, by Beyotime (1 mentions) Trans blot turbo, by Bio-Rad (1 mentions) Hrp labeled goat anti rabbit igg h l, by Beyotime (1 mentions)

Spelling variants of this product

Primary antibody dilution buffer (77 citations) QuickBlock (34 citations) QuickBlock buffer (23 citations) Antibody dilution buffer (13 citations) QuickBlock Western (13 citations) QuickBlock™ Primary Antibody Dilution Buffer (11 citations) Dilution buffer (9 citations) QuickBlock western buffer (7 citations) Primary antibody dilution buffer for western blot (2 citations) Western Primary Antibody Dilution Buffer (2 citations)

7 protocols using quickblock primary antibody dilution buffer for western blot

Revealing Protein Profiles via SDS-PAGE and Western Blot

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Proteins
were separated by SDS-PAGE: protein samples were loaded onto 4–20%
gradient sodium dodecyl sulfate polyacrylamide preformed gels (FuturePAGE,
ET12420Gel, Nanjing ACE Biotechnology Co., Ltd., Nanjing, China),
and the gels were run in a MOPS-SDS running buffer (FuturePAGE, F00004Gel,
Nanjing ACE Biotechnology Co., Ltd., Nanjing, China). Western blot
was performed by transferring the proteins from the gels to a 0.45
μm poly(vinylidene difluoride) (PVDF) membrane (Millipore, Bedford,
MA). PageRuler prestained protein ladder (Thermo Fisher Scientific
Baltics UAB, Vilnius, Lithuania) was used as a protein marker. The
PVDF membrane was then blocked with QuickBlock blocking buffer for
Western Blot (Beyotime, Songjiang District, Shanghai) on a shaker
at room temperature for 30 min. Subsequently, the membrane was incubated
with 1:20 diluted sera (QuickBlock primary antibody dilution buffer
for Western Blot, Beyotime, Songjiang District, Shanghai) at 4 °C
overnight. After washing three times, the membrane was incubated with
a 1:500-diluted HRP-conjugated antihuman IgE (epsilon) antibody (Sigma,
St. Louis, MO). After that, the optical signals were visualized by
an ECL substrate (Beijing Aoqiang Biotechnology CO., Ltd., Beijing,
China) using a ChemiDoc Touch chemiluminescence imaging system (Bio-Rad,
Hercules, CA).

Hu J., Zhu L.P., Wang R.Q., Zhu L., Chen F., Hou Y., Ni K., Deng S., Liu S., Ying W., Sun J.L., Li H, & Jin T. (2024). Identification, Characterization, Cloning, and Cross-Reactivity of Zan b 2, a Novel Pepper Allergen of 11S Legumin. Journal of Agricultural and Food Chemistry, 72(14), 8189-8199.

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Quantifying Antioxidant Proteins and Oxidative Stress Markers

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The kidney tissue (weighed 0.1 mg) was homogenized in 500 μL RIPA lysis buffer (P0013B, Beyotime Biotechnology, China). Total protein was extracted from the supernatants after centrifugation at 11,000 × g for 20 min at 4°C. The protein content was determined using Bradford protein assay. Proteins were separated by polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride membrane (IPVH00010, EMD Millipore, Billerica, MA, USA). The membrane was blocked in 5% bovine serum albumin at room temperature for 1 hour and probed overnight at 4°C with primary antibodies specific for GPX4 (ab125066, Abcam, Cambridge, UK, 1 : 1000), superoxide dismutase 2 (SOD2) (13141S,CST, Boston, MA, 1 : 1000), and 4-hydroxynonenal (4HNE) (ab46545, Abcam, Cambridge, UK, 1 : 2000). Primary antibodies were diluted by QuickBlock™ Primary Antibody Dilution Buffer for Western Blot (P0256, Beyotime Biotechnology, China). The blots were then incubated at room temperature for 2 hours with goat anti-rabbit or anti-mouse secondary antibodies (both diluted at 1 : 5000), as appropriate. Protein bands were visualized and scanned using a ChemiDoc XRS+ Imaging System (Bio-Rad, Inc., USA). Blot images were acquired for quantification using a digital imager and analyzed with imaging software.

Zhou F., Yang Y., Luo L., Chen Y., Luo Q, & Chen J. (2020). Impact of Prolonged Mechanical Ventilation on Ferroptosis in Renal Ischemia/Reperfusion Injury in Rats. BioMed Research International, 2020, 6097516.

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Fab Binding Capacity Quantification

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The binding capacity of each Fab to knob proteins was determined by western blot. Ten micrograms of purified knob proteins were preheated (100 °C for 10 min) and then separated and analyzed by electrophoresis on 10% SDS-PAGE under denaturing and reducing conditions, and transferred onto polyvinylidene fluoride membranes. The membranes were then blocked with QuickBlock Blocking Buffer for Western Blot (Beyotime Biotechnology, Shanghai, China) overnight. After washing again with PBS, Fab (5 μg/mL) in QuickBlock Primary Antibody Dilution Buffer for Western Blot (Beyotime Biotechnology, Shanghai, China) was added and incubated at 25 °C for 2 h. After washing thrice with 0.05% PBST, bound Fab was detected by incubation with a 1:5000 dilution of HRP-conjugated anti-HA tag mouse mAb (Abbkin, Redlands, CA, USA) in QuickBlock Secondary Antibody Dilution Buffer for Western Blot (Beyotime Biotechnology, Shanghai, China) at 25 °C for 1 h, and washed thrice in 0.05% PBST. The blot was developed by the ECL Detection System (UVP Chemstudio, Analytik Jena AG, Thuringia, Germany).

Liu Z., Tian X., Liu W., Xian Y., Chen W., Chen H, & Zhou R. (2019). Development of two antigen-binding fragments to a conserved linear epitope of human adenovirus and their application in immunofluorescence. PLoS ONE, 14(6), e0219091.

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Tissue Distribution of On-LECT2 in Tilapia

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The tissue distribution and relative expression of On-LECT2 in healthy tilapia at the protein level was assessed via Western blotting. Total protein from muscle, skin, brain, head kidney, liver and spleen tissues were isolated using a protein extraction kit (Solarbio, Beijing, China). Each sample containing 20 μg of total protein was loaded on 12% SDS-PAGE and transferred to a PVDF membrane (Millipore). The samples were blocked with QuickBlock™ Blocking Buffer for Western Blot (Beyotime, Shanghai, China) overnight at 4°C and then incubated with anti-LECT2 (PAF541Mu01, Cloud-Clone Corp, Wuhan, China) and anti-β-actin (AF5003, Beyotime, Shanghai, China) as primary antibodies. The solutions were diluted at a ratio of 1:1000 in QuickBlock™ Primary Antibody Dilution Buffer for Western Blot (Beyotime, Shanghai, China) for 2 h at room temperature. Afterwards, the membranes were washed three times in TBST and incubated with HRP-labelled goat anti-rabbit IgG (H+L) (Beyotime, Shanghai, China) at room temperature for 1 h. Antigen–antibody complexes were detected via the enhanced chemiluminescence method (P0018S, Beyotime, Shanghai, China).

Li Q., Zhang Z., Fan W., Huang Y., Niu J., Luo G., Liu X., Huang Y, & Jian J. (2021). LECT2 Protects Nile Tilapia (Oreochromis niloticus) Against Streptococcus agalatiae Infection. Frontiers in Immunology, 12, 667781.

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Western Blot Antibody Dilution Protocol

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HRP Goat Anti-Rabbit IgG (H+L) (AS014), AUF1 Rabbit pAb (A15679), GAPDH Rabbit mAb (A19056), and LCK Rabbit pAb (A2177) were bought from Abclonal. Primary antibodies and secondary antibodies were diluted in QuickBlock™ Primary Antibody Dilution Buffer for Western Blot (Beyotime) and QuickBlock™ Secondary Antibody Dilution Buffer for Western Blot respectively (Beyotime).

Zhang T., Liao D, & Hu Y. (2023). Cuproptosis-related lncRNAs forecast the prognosis of acute myeloid leukemia. Translational Cancer Research, 12(5), 1175-1195.

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Western Blot Analysis of Recombinant GST-Fusion Protein

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The purified rGST-On-CRP was loaded on 12% SDS-PAGE and transferred to a PVDF membrane (IPVH00010, Merck, Darmstadt, Germany) and then blocked with QuickBlock™ Blocking Buffer for Western Blot (Beyotime, Shanghai, China) at 25 °C for 15 min. The membrane was then incubated with anti-GST-tag antibodies (AG768, Beyotime, Shanghai, China), diluted at a ratio of 1:1000 in QuickBlock™ Primary Antibody Dilution Buffer for Western Blot (Beyotime, Shanghai, China) and incubated at 25 °C for 1 h.
Then the membrane was washed three times in TBST (TBS + 0.1% Tween 20) and incubated with secondary antibody HRP-labeled goat anti-rabbit IgG (H + L) (A0208, Beyotime, Shanghai, China), diluted at a ratio of 1:2000 in QuickBlock™ Secondary Antibody Dilution Buffer for Western Blot (Beyotime, Shanghai, China) at 25 °C for 30 min. Finally, the membrane was washed three times in TBST, and the antigen–antibody complexes were detected via the enhanced chemiluminescence method (P0018S, Beyotime, Shanghai, China).

Li Q., Jiang B., Zhang Z., Huang Y., Xu Z., Chen X., Cai J., Huang Y, & Jian J. (2022). CRP Involved in Nile Tilapia (Oreochromis niloticus) against Bacterial Infection. Biology, 11(8), 1149.

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Protein Acetylation Analysis by Western Blot

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Protein concentrations were determined by Bradford Protein Assay (P0006C, Beyotime, Shanghai, China). For Western blot, purified PykF and its variants were fractionated on a 4–20% SDS-PAGE gel and transferred to the PVDF membrane by Trans-Blot Turbo (Bio-rad, USA). The samples were blocked with QuickBlock™ Blocking Buffer for Western blot (P0239, Beyotime, Shanghai, China) overnight at 4°C and then incubated with the horseradish peroxidase (HRP)-conjugated acetyl lysine antibody (9441S, Cell Signaling Technology, USA) as primary antibodies. The solutions were diluted at a ratio of 1: 2000 in QuickBlock™ Primary Antibody Dilution Buffer for Western blot (P0239, Beyotime, Shanghai, China) for 2 h at room temperature. Afterward, the membranes were washed three times in TBST and incubated with HRP-labeled goat anti-rabbit IgG (H+L) (A0208, Beyotime, Shanghai, China) at room temperature for 1 h. Antigen–antibody complexes were detected via the enhanced chemiluminescence method (P0018, Beyotime, Shanghai, China).

Xu Z., Wang L., Wang X., Wan M., Tang M, & Ding Y. (2022). Characterizing the Effect of the Lysine Deacetylation Modification on Enzyme Activity of Pyruvate Kinase I and Pathogenicity of Vibrio alginolyticus. Frontiers in Veterinary Science, 9, 877067.

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