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As014

Manufactured by ABclonal
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Sourced in China, United States

AS014 is a laboratory pipette designed for precise liquid handling. It is a single-channel pipette capable of accurately dispensing volumes ranging from 0.1 to 10 microliters. The pipette features an ergonomic design and accurate volume adjustment to ensure consistent and reliable results during laboratory experiments and procedures.

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Lab products found in correlation

As003, by ABclonal (25 mentions) Ac026, by ABclonal (12 mentions) Pvdf membrane, by Merck Group (12 mentions) A19056, by ABclonal (6 mentions) Ripa lysis buffer, by Beyotime (5 mentions) Bca protein assay kit, by Beyotime (4 mentions) Ac001, by ABclonal (4 mentions) Polyvinylidene fluoride membrane, by Merck Group (4 mentions) Bca assay kit, by Beyotime (3 mentions) Gapdh, by ABclonal (3 mentions) Chemidoc xrs plus luminescent image analyzer, by Bio-Rad (3 mentions) Pvdf membrane, by Roche (3 mentions) β actin, by Wuhan Servicebio Technology (2 mentions) P0013, by Beyotime (2 mentions) Ab1791, by Abcam (2 mentions) Ab231303, by Abcam (2 mentions) Ab94744, by Abcam (2 mentions) A2708, by ABclonal (2 mentions) A5786, by ABclonal (2 mentions) A2851, by ABclonal (2 mentions)

85 protocols using as014

1

Western Blot Analysis of Sox18 and β-Catenin

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The protein was extracted using a Total Protein Extraction Kit (Beyotime, Shanghai, China), and the concentration was measured using a BCA Protein Assay Kit (Beyotime, Shanghai, China). The proteins were separated and transferred to PVDF membranes. The PVDF membranes were probed with the primary antibodies and the secondary antibodies. Finally, the expression of protein was measured using the ECL Western Blot kit (Biosharp, Hefei, China) and analyzed by a ChemiDocTM Touch Imaging System (Bio-Rad, CAL, USA).
The primary antibodies were anti-Sox18 (1:1000, DF8720, Affinity Biosciences, Changzhou, China), anti-β-catenin (1:1000, AF0069, Beyotime, Shanghai, China) and anti-GAPDH (1:3000, T0004, Affinity Bio-sciences, Changzhou, China). The secondary antibodies were HRP goat anti-rabbit IgG (1:1000, AS014, ABclonal, Wuhan, China) and HRP goat anti-mouse IgG (1:1000, AS014, ABclonal, Wuhan, China).
Hu T., Lv X., Getachew T., Mwacharo J.M., Haile A., Quan K., Li Y., Wang S, & Sun W. (2022). Effect of Sox18 on the Induction Ability of Dermal Papilla Cells in Hu Sheep. Biology, 12(1), 65.
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2

Western Blot Analysis of HMOX1 and LC3 in Nodosin-Treated SW480 Cells

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The nodosin-treated SW480 cells were lysed and proteins were extracted from them using lysis buffer (P0013, Beyotime, China). The protein extracts were processed by SDS polyacrylamide gel electrophoresis and blotted on PVDF membranes. Western blotting (WB) was performed using the following primary antibodies: HMOX1 (Rabbit, 1:3,000, 10701-1-AP, proteintech, China), LC3 (Rabbit, 1:2,000, 14600-1-AP, proteintech, China), β-Actin (Rabbit, 1:2,000, GB11001, Servicebio, China). After the primary antibody was incubated and washed, the immunohy bridization signal was captured using HRP-conjugated secondary antibody. The secondary antibodies were listed as follows: HRP Goat Anti-Rabbit IgG (H + L) (1:4,000, AS014, ABclonal, China). The development was then performed using ECL solution (P0018S, Beyotime, China).
Fan H., Hao X., Gao Y., Yang J., Liu A., Su Y, & Xia Y. (2022). Nodosin Exerts an Anti-Colorectal Cancer Effect by Inhibiting Proliferation and Triggering Complex Cell Death in Vitro and in Vivo. Frontiers in Pharmacology, 13, 943272.
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3

Protein Extraction and Western Blot Analysis

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Proteins were extracted from freshly frozen tissues. Briefly, the tissues were homogenized and lysed with radio immunoprecipitation lysis buffer (Solarbio, R0010, Beijing, China), which has 100 mg/ml of phenylmethanesulfonyl fluoride (ADOOQ, A11901, California, USA) and 1 mg/ml of Aprotinin (ADOOQ, A14263). The lysate was collected and centrifugated for liquid supernatant. A total of 50 μg protein samples were separated with 10% SDS-PAGE (Solarbio, P1040) and transferred onto polyvinylidene fluoride (Millipore, HVLP04700, Arklow, Ireland) membranes. Membranes were blocked at room temperature with 5% skim milk (ACMEC, AC11037, Shanghai, China) powder in TBS (Quartett, 402000192, Berlin, Germany) for 1 h. Membranes were then incubated overnight at 4 °C with primary antibodies against the following proteins: Acyl-CoA synthetase long-chain familymember 4 (ACSL4) (1: 1000, ABclonal, A6826, Wuhan, China), transferrin receptor (TFRC) (1: 1000, ABclonal, A5865, Wuhan, China). Secondary antibodies (1: 1000, ABclonal, AS014, Wuhan, China) were incubated for 1 h at 37 °C and washed three times with PBST (PBS+0.1% Tween 20) (Biomed, PA202, Beijing, China). The protein bands were visualized by Molecular Imager Gel Doc XR System (Bio-Rad, Hertfordshire, UK) and quantified by densitometric analysis using an image analyzer (NIH Image J software, Bethesda, USA).
Yu X., Ma X., Lyu J., Jiang N., Lu Y., Liao Y., Wang K, & Yu W. (2022). Ferroptosis involved in sevoflurane-aggravated young rats brain injury induced by liver transplantation. Neuroreport, 33(16), 705-713.
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4

Histone Acetylation Analysis Protocol

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The ground fine powder of each time point was mixed with extraction buffer containing 100 mM Tris, pH 8.0, 1.5 M β-mercaptoethanol, 4% SDS, 15% glycerol, 2 mM EDTA, 0.005% bromophenol blue, 2 μM/mL Trichostatin A (TSA) and vortexed for 30 s. The mixed solution was boiled for 5 min at 99 °C and vortexed 30 s. The supernatants were transferred into a new collection tube after centrifugation at 15,000 g for 10 min at RT. Proteins were loaded into 12% sodium dodecylsulfate-polyacryamide gel electrophoresis (SDS-PAGE) for protein separation and transferred to a polyvinylidene fluoride (PVDF) membrane, and probed using anti-H3 (ab1791) and anti-H3K18ac (ab1191) from Abcam, anti-H3K23ac (#07–355), anti-H3AC (#06–599), anti-H4K5ac (#07–327), anti-H4K8ac (#07–328), anti-H4K12ac (#07–595) and anti-H4K16ac (#07–329) from Millipore at a dilution of 1:2000 (v/v) in 5% skim-milk TBST (10 mM Tris-HCl, pH 75, 150 mM NaCl, and 0.1% Tween 20) for 1 h at RT after incubation with blocking buffer for 1 h at RT. The membranes were washed three times with TBST for 5 min each. Next, membranes were incubated with secondary antibody (AS014, ABclonal, Inc) in 5% skim-milk-TBST for 1 h at RT. Signals in membranes were detected using the BLT Gel View 6000 Plus machine (Biolight Biotec Co., Ltd.).
Guo J., Ma Z., Deng C., Ding J, & Chang Y. (2022). A comprehensive dynamic immune acetylproteomics profiling induced by Puccinia polysora in maize. BMC Plant Biology, 22, 610.
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5

Western Blot Analysis of Epithelial-Mesenchymal Transition

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Radioimmunoprecipitation assay (RIPA) was utilized to extract total cellular protein content, after which a BCA assay (Beyotime, Shanghai, China) was used for protein quantification. Samples were then electrophoretically separated and transferred onto PVDF membranes (Millipore, MA, USA). Following blocking for 2 h with 5% nonfat milk, blots were examined with proper primary and secondary antibodies. The bands of protein were then discovered via enhanced chemiluminescence (ECL) and analyzed with ImageJ (NIH, MD, USA). Antibodies used in this study were as follows: E-cadherin (abcam, ab231303, 1 : 1000), N-cadherin (abcam, ab98952, 1 : 1000), Vimentin (abcam, ab92547, 1 : 1000), Slug (abcam, ab27568, 1 : 1000), Snail (abcam, ab216347, 1 : 1000), HRP Goat Anti-Rabbit IgG (H+L) (ABclonal, AS014, 1 : 4000), HRP Goat Anti-Mouse IgG (H+L) (ABclonal, AS003, 1 : 5000).
Zhang Y., Yuan J., Guo M., Xiang R., Wang X., Xie T., Zhuang X., Li Q, & Lai Q. (2022). miR-657 Targets SRCIN1 via the Slug Pathway to Promote NSCLC Tumor Growth and EMT Induction. Disease Markers, 2022, 4842454.
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6

Sperm Protein Extraction and Western Blot Analysis

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Sperm total protein was extracted with sodium dodecyl sulfate (SDS) sample buffer. The total protein (20 µg) of each sample was separated by 10% SDS-PAGE gel (EC0023-B, Sparkjade, Jinan, China) and transferred onto a polyvinylidene fluoride (PVDF) membrane. The non-specific bindings of PVDF membranes were blocked with 5% (m/v) bovine serum albumin diluted with TBST (1% TBS, 0.1% Tween 20). These membranes were then immunoblotted with diluted primary antibodies against PRODH (22980-1-AP, 1:3000, Proteintech, Wuhan, China), Caspase3(A2156, 1:1000, AB clonal, Wuhan, China), Caspase9 (A0281, 1:1000, AB clonal, Wuhan, China) or tubulin (AC008, 1:1000, AB clonal, Wuhan, China) in TBST solution (1:1000 final dilution) and incubated at 4 °C for 12 h. Subsequently, membranes were washed with TBST solution and incubated with secondary antibodies (AS014, 1:1000, AB clonal, Wuhan, China) for 2 h, and then washed with TBST solution for three times. The ECL plus (ED0016-B, Sparkjade, Jinan, China) was used for detection and developed by gel imaging analyzer (Alpha, Fluor Chem Q, Shanghai, China).
Zhang W., Min L., Li Y., Lang Y., Hoque S.A., Adetunji A.O, & Zhu Z. (2022). Beneficial Effect of Proline Supplementation on Goat Spermatozoa Quality during Cryopreservation. Animals : an Open Access Journal from MDPI, 12(19), 2626.
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7

Western Blot Analysis of Osteoblast Markers

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Total protein of MC3T3 cells were harvested with RIPA lysis buffer and quantified with a BCA assay kit (Beyotime). Proteins were separated on 12% SDS-polyacrylamide gels and transferred to polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). These membranes were blocked with 5% bull serum albumin in Tris buffered saline with Tween 20 and incubated with the following primary antibodies at 4 °C overnight: cyclin D1 (1:500, A2708, ABclonal, Wuhan, China), SP7 (1:1000, ab94744, Abcam, Shanghai, China), OCN (1:650, A5786, ABclonal), Runx2 (1:500, A2851, ABclonal), P27Kip1 (1:500, A2692, ABclonal), and β-actin (1:100000, AC026, ABclonal). Horseradish peroxidase-conjugated anti-rabbit IgG (H + L) secondary antibodies were added (1:3000, AS014, ABclonal) and incubated at 25 °C for 1 h. The signals were detected using an ECL chemiluminescence kit (7Sea biotech, Shanghai, China) by Tanon 5200 (Tanon, Shanghai, China).
Yu W., Zheng Y., Li H., Lin H., Chen Z., Tian Y., Chen H., Zhang P., Xu X, & Shen Y. (2020). The Toll-like receptor ligand, CpG oligodeoxynucleotides, regulate proliferation and osteogenic differentiation of osteoblast. Journal of Orthopaedic Surgery and Research, 15, 327.
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8

Quantifying Tumor Protein Levels

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We extracted proteins from normal and adjacent tumor tissues and tested their concentrations using BCA protein kits (Boster, Wuhan, China). Target proteins were isolated with SDS-PAGE (10%), and transferred to nitrate fiber membrane (4 ℃, 270 mA, 1.5 h). Then we sealed the nitrate fiber membrane with 5% skim milk powder for 3 h. The membrane was incubated with rabbitanti-CASP3 antibody (1:2,000; cat. no. ab184787; Abcam) or mouse anti‑glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (1:5,000; cat. no. ab8245; Abcam) orrabbit anti-TP53 antibody (1:500; cat. no. M00001-4; Boster) orrabbit anti-β-actin antibody (1:500; cat. no. WL0002d; Wanleibio) at 4 ℃ over-night. Wash unbound protein 3 times with 5% TBST solution., and the membrane was incubated with HRP Goat Anti-Rabbit IgG (H+L) secondary antibody (1:4,000; AS014; Abclonal) or HRP Goat Anti-Mouse IgG (H+L) secondary antibody (1:4,000; E-AB-1001; ELAbscience) for 1 h. Following washing with TBST (3 times). ECL western blot detection kit (Affinity Biosciences, USA) was used to visualize the protein bands. β-actin was used as internal references for protein. The molecular weight and net optical density of target band were analyzed by Image J. analysis software, and the results represented the relative content of target protein.
Liu S., Zhao Y., Duan R., Wu Y., Chen X, & Li N. (2022). Identification of core genes associated with type 2 diabetes mellitus and gastric cancer by bioinformatics analysis. Annals of Translational Medicine, 10(5), 247.
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9

Quantification of AMPK Activation in C. elegans

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Pellets of 100,000 eggs obtained by bleaching, 20,000 L2s, 5000 L4s and 2000 3-day-old adults were collected and snap-frozen with liquid nitrogen. All worm samples were preserved in a –80 °C freezer before use. For western blot assays, all samples were thawed on ice and lysed in RIPA-mid (Beyotime, P0013C) with sonication. To avoid dephosphorylation of the AAK-2 (AMPKα) protein, a phosphatase inhibitor was added to the samples before sonication. The worm lysates were then centrifuged at 16,000× g for 10 min at 4 °C, and the supernatant was collected for protein concentration determination with a BCA protein assay. Forty micrograms of total protein per well was loaded. The membranes were incubated with primary antibodies anti-pAMPK (Cell Signaling, 2535S, 1:1000, diluted with 2% BSA) and anti-tubulin (Abcam, ab6161, 1:3000, diluted with 2% BSA) and with HRP goat-anti-rabbit (Abclonal, AS014, 1:10,000, diluted with 5% milk), and HRP goat-anti-rat (Beyotime, A0192, 1:5000, diluted with 5% milk) secondary antibodies.
Zhang R., Fang J., Qi T., Zhu S., Yao L., Fang G., Li Y., Zang X., Xu W., Hao W., Liu S., Yang D., Chen D., Yang J., Ma X, & Wu L. (2023). Maternal aging increases offspring adult body size via transmission of donut-shaped mitochondria. Cell Research, 33(11), 821-834.
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10

Immunohistochemical Analysis of CD31 and VWF

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Sections were incubated with an anti-CD31 antibody (dilution 1:100, A3181; ABclonal) or an anti-VWF antibody (dilution 1:100, A1335; ABclonal) overnight at 4°C followed by incubation with goat anti-rabbit IgG (H+L) horseradish peroxidase (HRP)-conjugated secondary antibody (dilution 1:200, AS014, ABclonal) for 2 h at room temperature. The reactions were visualized using a 3,3′-diaminobenzidine visualization kit (Fuzhou Maixin Biotech Co., Ltd.). Sections were counterstained with hematoxylin to visualize nuclei, stained with hematoxylin for 5–10 min at room temperature, and dehydrated. Sections were examined under a light microscope at a magnification of ×200. Brown staining indicated immunoreactive positive cells, and blue staining indicated the nuclei.
Chen M., Huang Z., Bi H., Pan X., He J., He L., He X., Du J., Zhou K., Wang L., Wang Q., Guo X, & Jin Z. (2019). Effects of bone marrow-derived mesenchymal stem cell transplantation on chronic obstructive pulmonary disease/obstructive sleep apnea overlap syndrome in rats. Molecular Medicine Reports, 20(5), 4665-4673.
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