Ab134146

Manufactured by Abcam

Sourced in United States

Ab134146 is a primary antibody provided by Abcam. It is designed for use in various immunological techniques.

Automatically generated - may contain errors

Lab products found in correlation

Ab76417, by Abcam (3 mentions) Ab32572, by Abcam (3 mentions) Ripa lysis buffer, by Merck Group (2 mentions) Ab54365, by Abcam (2 mentions) Ab6258, by Abcam (2 mentions) Ab6528, by Abcam (2 mentions) Ab32358, by Abcam (2 mentions) Ab51608, by Abcam (2 mentions) Hrp conjugated goat anti rabbit igg h l, by Abcam (2 mentions) Ab92547, by Abcam (2 mentions) Epr2423 2, by Abcam (1 mentions) Ab203201, by Abcam (1 mentions) Af555, by Thermo Fisher Scientific (1 mentions) Anti n cadherin, by Thermo Fisher Scientific (1 mentions) Anti e cadherin, by Abcam (1 mentions) Ab6160, by Abcam (1 mentions) Ab11512, by Abcam (1 mentions) Ab183991, by Abcam (1 mentions) Rcab002p f, by ReproCELL (1 mentions) Anti mcherry, by Rockland Immunochemicals (1 mentions)

11 protocols using ab134146

Murine and Human MSC Surface Marker Profiling

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For murine MSCs, non-permeabilized bone marrow cells collected from mice were incubated with the antibodies against MSCs cell surface markers including anti-CD45-APC, anti-Ter119-APC, anti-CD31-APC, and anti-LepR-biotin. For cell surface CD36, LRP5 and LRP6 detection, anti-CD36 (Biolegend, 102606, 1:200), anti-LRP5 (Abcam, ab203201, 1:200) or anti-LRP6 [EPR2423(2)] (Abcam, ab134146, 1:200) were added individually to the non-permeabilized cells followed by FITC-conjugated secondary antibody. The frequency of CD36-, LRP5-, or LRP6-positive cells in the total LepR⁺CD45^–CD31^–Ter119^- cell population was calculated.
To detect cell surface CD36, LRP5 and LRP6 expression in human MSCs, non-permeabilized human MSCs purchased from Lonza were incubated with individual antibodies: anti-CD36 (Biolegend, 102606, 1:200), anti-LRP5 (Abcam, ab203201, 1:200) or anti-LRP6 [EPR2423(2)] (Abcam, ab134146, 1:200) followed by FITC- and PE-conjugated secondary antibody. The frequency of CD36-, LRP5-, or LRP6-positive cells in the total MSCs was calculated. Flow cytometry analyses were performed using a LSR II flow cytometer.

Wang L., Chai Y., Li C., Liu H., Su W., Liu X., Yu B., Lei W., Yu B., Crane J.L., Cao X, & Wan M. (2018). Oxidized phospholipids are ligands for LRP6. Bone Research, 6, 22.

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Comprehensive Immunostaining Protocol

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The antibodies used were anti-α-tubulin (YL1/2; rat; ab6160; Abcam), anti–β-catenin (mouse; #610154; BD Transduction), anti-LRP6 (EPR2423(2); rabbit; ab134146; Abcam), anti-NANOG (rabbit; RCAB002P-F; Reprocell), anti-OCT3/4 (mouse; #611202; BD Transduction), anti-EOMES (rabbit; ab183991; Abcam), anti-GriA3 (mouse; MAB5416; Sigma-Aldrich), anti-GriA4 (rabbit; AB1508; Sigma-Aldrich), anti-GriK1 (rabbit; AGC-008; Alomone Laboratories), anti-GriK3 (rabbit; AGC-040; Alomone Laboratories), anti–N-cadherin (mouse; #33-3900; Thermo Fisher Scientific), anti–E-cadherin (DECMA-1; rat; ab11512; Abcam), anti-GFP (chicken; GFP-1020; Aves), anti-mCherry (goat; #200-101-379; Rockland), and AF488, AF555, or AF647-conjugated secondary antibodies (Thermo Fisher Scientific).

Junyent S., Reeves J., Gentleman E, & Habib S.J. (2021). Pluripotency state regulates cytoneme selectivity and self-organization of embryonic stem cells. The Journal of Cell Biology, 220(4), e202005095.

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Immunofluorescence Staining of GC Cells

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GC cells or frozen GC tissue sections were fixed for 15 min in 4% paraformaldehyde. After washing with PBS, the sections were blocked with a protein-blocking solution. A rabbit monoclonal antibody against LRP6 (Cat. no. ab134146, Abcam) or a goat polyclonal antibody that recognizes all isoforms of CMG2 (Cat. no. SAB2501374, Sigma-Aldrich) or rabbit monoclonal antibody against CD44 (Cat. no. BBA10, R&D) or CK18 (Cat. no. ab82254, Abcam) or H-KATPase (Cat. no. ab2866, Abcam) was added onto the slides. After incubation overnight at 4 °C, the sections were incubated with Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (Invitrogen, Carlsbad, CA) and/or Alexa Fluor® 647 donkey anti-goat IgG (H+L) at 37 °C for 30 min. Cell nuclei were then stained with Hoechst 33342. All samples were then analyzed by a confocal laser scanning microscopy.

Ji C., Yang L., Yi W., Xiang D., Wang Y., Zhou Z., Qian F., Ren Y., Cui W., Zhang X., Zhang P., Wang J.M., Cui Y, & Bian X. (2018). Capillary morphogenesis gene 2 maintains gastric cancer stem-like cell phenotype by activating a Wnt/β-catenin pathway. Oncogene, 37(29), 3953-3966.

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Automated Western Blotting of Wnt Signaling Proteins

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Automatic western blots were performed using a Wes automated system (ProteinSimple, California, USA). Purified recombinant proteins were used as calibration standards. Serial dilutions of both the sample and standard were used to determine the linear dynamic range of the assay. According to the ProteinSimple kit (SM-W004), the loading order was: ladder and Experimental protein samples; antibody Diluent II; antibody Diluent II and Primary Antibody; streptavidin-HRP and Secondary HRP Conjugate; luminol-Peroxide mix. SW software was used for data generation and analysis. Antibodies against p-LRP6 (ab226758, 1:50), LRP6 (ab134146, 1:10), p-GSK3b (ab68476, 1:100), GSK3b (ab185141, 1:100), β-catenin (ab16051, 1:50), LEF1 (ab137872, 1:10), and β-actin (ab6276, 1:100) were obtained from Abcam. Antibodies against p-β-catenin (4176, 1:100) were obtained from Cell Signaling Technology.

Sun J., Zhang H., Liu D., Liu W., Du J., Wen D., Li L., Zhang A., Jiang J, & Zeng L. (2023). CTGF promotes the repair and regeneration of alveoli after acute lung injury by promoting the proliferation of subpopulation of AEC2s. Respiratory Research, 24, 227.

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Protein Expression Profiling in WiT49 Cells

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The levels of HOXA11, β-catenin, receptor–related protein 6 (LRP6), phosphorylation-LRP6 (p-LRP6), E-cadherin (E-cad), N-cadherin (N-cad), Vimentin, HIF-1α, C/EBPβ, and β-actin protein expression were assessed in WiT49 cells. The total cellular proteins were isolated with RIPA lysis buffer (#R0278, Sigma), and a standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method was used to separate the proteins [19 ]. Next, the separated protein bands were transferred onto membranes that were subsequently blocked, and then cultured with primary antibodies against HOXA11 (#ab54365; 1:1000), β-catenin (#ab32572; 1:800), LRP6 (#ab134146; 1:1000), p-LRP6 (#ab76417; 1:800), E-cad (#ab6528; 1:800), N-cad (#ab6258; 1:800), Vimentin (#ab92547; 1:800), HIF-1α (#ab51608; 1:1000), C/EBPβ (#ab32358; 1:1000), and β-actin (#ab32572; 1:5000) (Abcam, Cambridge, MA, USA). Next, the membranes were then washed and incubated with HRP conjugated Goat Anti-Rabbit IgG H&L (1: 5000, #ab6721, Abcam). Finally, the immunostained bands were observed using a Tanon 6600 Luminescence imaging workstation (Tanon, China).

Zhu S., Zhou R., Tang X., Fu W, & Jia W. (2024). Hypoxia/inflammation-induced upregulation of HIF-1α and C/EBPβ promotes nephroblastoma cell EMT by improving HOXA11-AS transcription. Heliyon, 10(6), e27654.

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Quantifying AML Cell Signaling Proteins

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AML cells (MV4-11 and MOLM-13) were drugged for 24 h, followed by protein extraction using RIPA buffer (Cell Signaling Technology) supplemented with PMSF. Cell pellets were incubated in RIPA buffer with constant agitation for 30 min, followed by sonication. Protein lysate was quantified using Pierce™ BCA Protein Assay Kit (ThermoFischer Scientific). 30 µg of protein was loaded into a Mini-PROTEAN TGX Stain-Free Precast Gel (Bio-Rad) and ran at 120 V for 1–2 h. Blots were developed using a ChemiDoc Chemiluminescence Imaging System (Bio-Rad) or by the Odyssey CLX Fluorescence Imaging System (Li-COR). Antibodies used in the presented studies purchased from Cell Signaling Technology included total β-catenin (#8480S), phosphorylated β-catenin (#9561S), non-phosphorylated β-catenin (cat#8814S), and β-actin (#3700S). Antibodies purchased from Abcam include LRP6 (ab134146) and phosphorlated-LRP6 (ab76417).

Marr A.R., Halpin M., Corbin D.L., Asemelash Y., Sher S., Gordon B.K., Whipp E.C., Mitchell S., Harrington B.K., Orwick S., Benrashid S., Goettl V.M., Yildiz V., Mitchell A.D., Cahn O., Mims A.S., Larkin K.T., Long M., Blachly J., Woyach J.A., Lapalombella R, & Grieselhuber N.R. (2024). The multi-CDK inhibitor dinaciclib reverses bromo- and extra-terminal domain (BET) inhibitor resistance in acute myeloid leukemia via inhibition of Wnt/β-catenin signaling. Experimental Hematology & Oncology, 13, 27.

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Investigating Endometrial Cancer Cell Lines

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Human endometrial cancer cells AN3CA, HEC-1A, ECC-1, Ishikawa and HEK293T were obtained as gifts from Shanghai Cancer Institute. All these cells were cultured according to American Type Culture Collection (ATCC) instructions. Antibodies used in this study were against SMOC-2 (ab78068, Abcam) for western blotting (WB) and Immunohistochemistry (IHC). CD133 (372805, Biolegend), CD44 (103008, Biolegend) for Flow Cytometry. CD133 (ab216323, Abcam), CD44 (ab51037, Abcam) and PCNA (13110, Cell Signaling) for IHC. β-actin (M1210–2, Huabio), β-Tubulin (66240–1-Ig) and Lamin A/C (2032s, cell signaling) for western blotting (WB), β-catenin (ab32572, Abcam) for WB, immunofluorescence (IF). Wnt3a (ab28472, Abcam), Wnt10b (ab70816, Abcam), Frizzled3 (sc-68334, Santacruz), Frizzled6 (5158, cell signaling), Frizzled8 (sc-33503, Santacruz) Frizzled6 (5158, cell signaling), LRP1 (ab92544, Abcam), LRP6 (ab134146, Abcam), LRP7 (ab36121, Abcam), HA-tag (3724, cell signaling), Flag-tag (8146, cell signaling), and Myc-tag (2278, cell signaling) for CO-IP. Secondary antibodies were purchased from Jackson. TUNEL assay was purchased from Sigma (11684817910). Chemicals and biochemicalused were Paclitaxel (EY1353, Amquar), cisplatin (EY0024, Amquar), FGF (091608, PeproTech), EGF (0816AFC05, PeproTech), B27 (17504044, Gibco) and XAV-939 (S1180, Selleck).

Lu H., Ju D.D., Yang G.D., Zhu L.Y., Yang X.M., Li J., Song W.W., Wang J.H., Zhang C.C., Zhang Z.G, & Zhang R. (2018). Targeting cancer stem cell signature gene SMOC-2 Overcomes chemoresistance and inhibits cell proliferation of endometrial carcinoma. EBioMedicine, 40, 276-289.

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Western Blot Protein Expression Analysis

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Cells were lysed in the RIPA lysis buffer [20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% NP-40, 1% sodium deoxycholate and 1 mM AEBSF]. Equal amounts of samples were subjected to a Tris-Glycine Gel (Invitrogen, XP0012C) in a Mini Gel Tank (Invitrogen, A25977). The resolved proteins were transferred onto a nitrocellulose membrane (Amersham Protran 0.2 μm, 10600006) using a wet electroblotting system (Bio-Rad Mini Protean II Cell) followed by immunoblotting. 5% non-fat dry milk in 1× TBS-T (0.1% Tween-20) was used for blocking at room temperature for 1 h.
Primary antibodies were applied overnight at 4°C as follows: LRP6, rabbit, Abcam, ab134146, 1:1,000; Cyclin D1, rabbit, Abcam, ab16663, 1:2,500; alpha-Tubulin, mouse, Merck Millipore, CP06, 1:10000; HSP90, rabbit, Cell Signaling Technology, #4874S, 1:1,000; GAPDH, mouse, Santa Cruz biotechnology, sc-32233, 1:10000.
Secondary antibodies: Goat anti-mouse IgG (HRP), Abcam, ab97265, 1:10000; Goat anti-rabbit IgG (HRP) Abcam, ab6721, 1:10000.
Signals were detected by SuperSignal West Dura (Life Technologies, 34075) with an Optimax 2010 X-Ray Film Processor (PROTECT) or using the BioRad ChemiDoc MP Imaging System. The results were quantified using ImageJ, with one-way ANOVA as a statistical analysis.

Geng S., Paul F., Kowalczyk I., Raimundo S., Sporbert A., Mamo T.M, & Hammes A. (2023). Balancing WNT signalling in early forebrain development: The role of LRP4 as a modulator of LRP6 function. Frontiers in Cell and Developmental Biology, 11, 1173688.

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Investigating Wnt9a-Klotho-LRP6 Interactions

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The interaction of Klotho or LRP6 and Wnt9a in HKC‐8 cells was determined by co‐immunoprecipitation, as previously reported (Zhou et al., 2013). Cells were transfected with Wnt9a expression vector (pFlag‐Wnt9a) for 24 hours, and then co‐treated with recombinant human Klotho (5334‐KL; R&D Systems) for 12 hours. Cell lysates were immunoprecipitated overnight at 4°C with an antibody against Wnt9a (ab125957; Abcam) using protein A/G plus agarose (sc‐2003; Santa Cruz Biotechnology). After washing three times, the precipitated complexes were immunoblotted with anti‐Wnt9a (ab125957; Abcam), anti‐Klotho (AF1819; R&D Systems), and anti‐LRP6 (ab134146; Abcam) antibodies. Cell lysates were also tested by an antibody against GAPDH (RM2001, Ray Antibody Biotech).

Miao J., Huang J., Luo C., Ye H., Ling X., Wu Q., Shen W, & Zhou L. (2021). Klotho retards renal fibrosis through targeting mitochondrial dysfunction and cellular senescence in renal tubular cells. Physiological Reports, 9(2), e14696.

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Wnt Signaling Pathway Investigation

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IWP-4 (#HY-12879) and JW55 (#HY-13968) were purchased from MedChemExpress (New Jersey, USA). CCK8 kit (#CA1210), 4% paraformaldehyde (#P1110), hematoxylin–eosin staining kit (#G1120), Trizol(#R1100), BCA Protein Quantitative Kit (#PC0020), and 30% acrylamide (29:1) (#A1010) were purchased from Solarbio life sciences (Beijing, China). HiFiScript gDNA Removal RT MasterMix (#CW2020) and SuperRT One Step RT-PCR Kit (#CW0742) were purchased from cwbiosciences (Jiangsu, China). Protein loading buffer (#P0015L), protein pre-staining Marker(#P0069), and BeyoECL Star Luminescent Liquid (#P0018AS) were purchased from Beyotime (Shanghai, China). Cr(C011-2-1), BUN (C013-2-1), AST (C010-2-1), and ALT(C009-2–1), detection kits were purchased from Nanjing Jiancheng Bioengineering Institue (Nanjing, Chian). Antibody for Wnt5 (#55184-1-AP), β-Catenin (#51067-2-AP), ARF6 (#20225-1-AP), and N-Cadherin (#22018-1-AP), and GAPDH (#60004-1-Ig), HRP-conjugated Affinipure Goat Anti-Rabbit IgG(H + L) (#SA00001-2), and FITC-conjugated Affinipure Goat Anti-Rabbit IgG(H + L) (#SA00003-2) were purchased from Proteintech (Wuhan, China). Antibody for LRP5 (ab223203), LRP6 (ab134146), TCF-4 (ab130014), and LEF1 (ab137872) were purchased from abcam (Cambridge, UK).

Bian Y., Wang G., Zhou J., Yin G., Liu T., Liang L., Yang X., Zhang W., Ni K., Tang D, & Yu Y. (2022). Astragalus membranaceus (Huangqi) and Rhizoma curcumae (Ezhu) decoction suppresses colorectal cancer via downregulation of Wnt5/β-Catenin signal. Chinese Medicine, 17, 11.

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