Acid phosphatase assay kit

Manufactured by Beyotime

Sourced in China

The Acid Phosphatase Assay Kit is a laboratory tool used to measure the activity of the enzyme acid phosphatase. This kit provides a standardized method for quantifying the levels of acid phosphatase in a given sample.

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Lab products found in correlation

Chloroquine phosphate, by Merck Group (1 mentions) Alexa fluor 647 labeled goat anti rabbit igg h l, by Beyotime (1 mentions) Succinic anhydride, by Adamas (1 mentions) B pei, by Adamas (1 mentions) Rabbit anti lc3b, by Proteintech (1 mentions) Rabbit anti cd44, by Beyotime (1 mentions) Anti p gp, by Proteintech (1 mentions) Ad gfp lc3b, by Beyotime (1 mentions) Anti p62, by Proteintech (1 mentions) Anti ki67, by Beyotime (1 mentions) Tunel apoptosis assay kit, by Beyotime (1 mentions) Lyso tracker red, by Beyotime (1 mentions) Lysis buffer, by Beyotime (1 mentions)

6 protocols using acid phosphatase assay kit

Succinic anhydride-conjugated PEI for P-glycoprotein silencing

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Succinic anhydride, b-PEI (MW 1,800), b-PEI (MW 250,000), 2-(7-Azabenzotriazol-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate (HATU), N-ethyldiisopropylamine (DPIEA) et.al was purchased from Adamas-beta (Shanghai, China). All of solvent were purchased from Macklin (Shanghai, China). Chloroquine Phosphate was obtained from Sigma-Aldrich (St. Louis, MO, USA). Antibodies used for Western Blotting and immunofluorescence including rabbit anti-LC3B, anti-p62, anti-Pgp, goat anti-rabbit IgG (H + L) (HRP, Cora Lite 594) were obtained from Proteintech (Wuhan, China). Alexa fluor 647-labeled goat anti-rabbit IgG (H + L), rabbit anti-CD44, anti-ki67, Ad-GFP-LC3B, Lyso-Tracker Red, TUNEL Apoptosis Assay Kit, acid phosphatase assay kit et.al were purchased from Beyotime (Shanghai, China). All other reagents for western blotting and gel electrophoresis were obtained from Solarbio (Beijing, China).
Targeting human P-gp siRNA sequences:
Sense: 5’-AAGAAGGAAAAGAAACCAACUdTdT-3’;
Anti-sense: 5’-AGUUGGUUUCUUUUCCUUCUUdTdT-3’.
All of siRNA were obtained by GenePharma Co. Ltd. (Shanghai, China).

Wang C., Li Z., Xu P., Xu L., Han S, & Sun Y. (2022). Combination of polythyleneimine regulating autophagy prodrug and Mdr1 siRNA for tumor multidrug resistance. Journal of Nanobiotechnology, 20, 476.

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Evaluation of Macrophage Immune Response

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Tongue sole were administered with or without pCsBAFF and pCN3, and at 7 days post-plasmid administration, head kidneys were removed from the fish and washed with PBS. Head kidney macrophage (HKM) preparation and respiratory burst analysis were performed as reported elsewhere [32 (link)]. Acid phosphatase activity was determined using the Acid Phosphatase Assay Kit (Beyotime Institute of Biotechnology, Beijing, China). All experiments were performed independently three times.

Sun Y, & Sun L. (2015). CsBAFF, a Teleost B Cell Activating Factor, Promotes Pathogen-Induced Innate Immunity and Vaccine-Induced Adaptive Immunity. PLoS ONE, 10(8), e0136015.

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Acid Phosphatase Activity Quantification

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The cells were lysed with lysis buffer (Beyotime; P0013J), and after centrifugation, the supernatant was obtained for phosphatase activity detection using an acid phosphatase assay kit (Beyotime, P0326). The test consisted of a control along with 4, 8, 16, 24, 32, and 40 μL of standard product and 40 μL of sample. After directly incubating for 5–10 min at 37°C, 160 μL stop solution was added to terminate the reaction. Absorbance was measured at 405 nm, and acid phosphatase activity was calculated according to the definition of enzymatic activity.

Zuo S.Q., Li C., Liu Y.L., Tan Y.H., Wan X., Xu T., Li Q., Wang L., Wu Y.L., Deng F.M, & Tang B. (2021). Cordycepin inhibits cell senescence by ameliorating lysosomal dysfunction and inducing autophagy through the AMPK and mTOR–p70S6K pathway. FEBS Open Bio, 11(10), 2705-2714.

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Respiratory Burst and Acid Phosphatase Assays

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For respiratory burst assay, rCsMAP34, rTrx, or HBSS (control) was added to PBL in a 96-well tissue culture plate (~10⁵ cells/well) to a final concentration of 80 μg/ml. The plate was incubated at 22 °C for 1 h, 2 h, or 4 h. After incubation, the cells were determined for respiratory burst as reported previously61 (link). For acid phosphatase activity assay, the above cells were lysed by adding 100 μl of 1% Triton X-100 to each well and incubation at 4 °C for 20 min. After incubation, acid phosphatase activity was determined using Acid Phosphatase Assay Kit (Beyotime, Beijing, China) according to manufacturer’s instruction.

Li M.F., Li J, & Sun L. (2016). CsMAP34, a teleost MAP with dual role: A promoter of MASP-assisted complement activation and a regulator of immune cell activity. Scientific Reports, 6, 39287.

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Respiratory Burst and Acid Phosphatase Activity in Macrophages upon rSil Treatment

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rSil was added to HKM in a 96-well tissue culture plate (∼10⁵ cells/well) to a final concentration of 10 µM, 20 µM, or 30 µM. The same volume of PBS was added to the control cells. The plate was incubated at 22°C for 1 h. S iniae SF1 was cultured as above and resuspended in L-15 to 10⁷ CFU/ml. One hundred microliters of S. iniae was added to each well of HKM. The plate was incubated at 22°C for 3 h. After incubation, the cells were determined for respiratory burst as reported previously [43] and for acid phosphatase activity as follows: the cells were lysed by adding 100 µl of 1% Triton X-100 to each well and incubation at 4°C for 20 min. After incubation, acid phosphatase activity was determined using Acid Phosphatase Assay Kit (Beyotime, Beijing, China) according to manufacturer's instruction.

Li M.F., Zhang B.C., Li J, & Sun L. (2014). Sil: A Streptococcus iniae Bacteriocin with Dual Role as an Antimicrobial and an Immunomodulator That Inhibits Innate Immune Response and Promotes S. iniae Infection. PLoS ONE, 9(4), e96222.

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Quantifying Bacillus cereus Infection

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IEC‐6 cells were infected with B. cereus at an MOI of 40 under 0.5 µg mL⁻¹ ciprfloxacin treatment at 37 °C for 8 h. The activity of ACP (U mL⁻¹) from cultural supernatants was detected by an Acid Phosphatase Assay Kit (Beyotime) though quantification of the consumption of phosphatase chromogenic substrates (Para‐nitrophenyl phosphate, pNPP), according to the manufacturer's instruction.

Liu X., Liu F., Ding S., Shen J, & Zhu K. (2020). Sublethal Levels of Antibiotics Promote Bacterial Persistence in Epithelial Cells. Advanced Science, 7(18), 1900840.

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