Ab6000

Manufactured by Merck Group

Sourced in United States

The AB6000 is a versatile laboratory equipment designed for analysis and data processing tasks. It features a high-performance processor, ample storage capacity, and advanced data management capabilities. The AB6000 is suitable for a wide range of laboratory applications, providing reliable and efficient performance.

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Lab products found in correlation

Ab1791, by Abcam (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Inverted fluorescence microscope, by Thermo Fisher Scientific (1 mentions) Pipettor, by Eppendorf (1 mentions) Ab11575, by Abcam (1 mentions) A5441, by Merck Group (1 mentions) B2531, by Merck Group (1 mentions) V8031, by Promega (1 mentions) Prb 236p, by Fortrea (1 mentions) Smi 94r, by Fortrea (1 mentions) Anti actin, by Merck Group (1 mentions) Anti mmp 1, by Merck Group (1 mentions) Multi gauge software, by Fujifilm (1 mentions) Trans blot turbo transfer system, by Bio-Rad (1 mentions) Anti app clone 22c11, by Merck Group (1 mentions) Ab947, by Merck Group (1 mentions) Ab180711, by Abcam (1 mentions)

5 protocols using ab6000

ChIP-seq Protocol for Epigenetic Marks

Cited 1 time

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ChIP was performed as previously described (Schwartz et al. 2012 (link); Davidovich et al. 2013 (link)) with the exceptions noted in the Supplemental Material. For immunoprecipitation, 5–25 µg of solubilized chromatin was used with 2 µg of α-RNA Pol II antibody (EMD Millipore, catalog no. 05-623), α-H3 (Abcam, ab-1791), α-H3K4me2/3 (Abcam, ab-6000), α-H3K27me3 (EMD Millipore, 07-449), or 4 µg of α-GABPα (Santa Cruz Biotechnology, H-180 sc-22810) and nutated overnight at 4°C.
All replicates reported in this study represent independent biological samples.

Stern J.L., Theodorescu D., Vogelstein B., Papadopoulos N, & Cech T.R. (2015). Mutation of the TERT promoter, switch to active chromatin, and monoallelic TERT expression in multiple cancers. Genes & Development, 29(21), 2219-2224.

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Quantitative Analysis of MMP-2, TIMP-3 and GAPDH

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MMP-2 monoclonal antibody, TIMP-3 and GAPDH monoclonal antibody (cat. nos. 04-1048, AB6000 and MABS819 all from EMD Millipore, Billerica, MA, USA), TRIzol, immunohistochemistry and reverse transcription kit (all from Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), inverted fluorescence microscope (Thermo Fisher Scientific, Inc.), sheep anti-rabbit IgG secondary antibody (cat. no. AP510, EMD Millipore, Billerica, MA, USA), pipettor (Eppendorf, Hamburg, Germany), polymerase chain reaction (PCR) instrument (ABI USA, Vernon, CA, USA), and an ultraviolet imaging system (Biometra GmbH, Göttingen, Germany) were used in the present study. The origin and batch number of other relevant instruments and materials are specified in corresponding parts.

Liu Z., Ren Y, & Zhu F. (2018). Expression of MMP-2 and TIMP-3 with incidence and prognosis of giant-cell tumor of the bone. Oncology Letters, 16(1), 721-726.

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Antibody Panel for Myelin and Cell Signaling Analysis

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The following primary antibodies were used for immunostaining or Western blot analysis: anti-mouse myelin basic protein (MBP; 1:300, #SMI-94R [Covance]), anti-rabbit Krox-20 (1:500, #PRB-236P [Covance]), anti-mouse BrdU (1:1000, #B2531 [Sigma]), anti-rabbit phospho-Akt (1:1000, #4051 [Cell Signaling Technology]), anti-rabbit phospho-Erk1/2 (1:5000, #V8031 [Promega]), anti-rabbit total-Akt (1:1000, #9272 [Cell Signaling Technology]), anti-rabbit total Erk1/2 (1:5000, #V1141 [Promega]), anti-rabbit phospho-ErbB3 (1:500, #4791 [Cell signaling]), anti-rabbit laminin (1:1000, #ab11575 [Abcam]), anti-mouse β-actin (1:5000, #A5441 [Sigma]), and anti-rabbit TIMP-3 (1:1000, #AB6000 [Millipore]).

Kim J., Elias A., Lee T., Maurel P, & Kim H.A. (2017). Tissue Inhibitor of Metalloproteinase-3 Promotes Schwann Cell Myelination. ASN NEURO, 9(6), 1759091417745425.

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Immunoblotting for Protein Validation

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A number of selected proteins that were detected as regulated in the analysis were further validated by immunoblotting (TIMP-3, TIMP-1, SPARC, MIF, APP). Supernatants from TIMP-3/HEK or HEK293 cells were loaded onto an acrylamide gel for electrophoresis. After electrophoretic separation, proteins were blotted onto a PVDF membrane using the Trans-Blot Turbo transfer system (Biorad) and detected by the following antibodies: anti-TIMP-3 (AB6000, Millipore), anti-TIMP-1 (generated as previously described^{65 (link)}), anti-TIMP-2 (generated as previously described^{66 (link)}), anti-SPARC (number 5031, Thermo Fisher), anti-MIF (clone FL-115, Santa Cruz) anti-APP (clone 22C11, Millipore), anti-MMP-1 (clone 2A7.2, Millipore) anti-actin (A5316, Sigma Aldrich).
Bands corresponding to each protein were quantified by using Multi Gauge software (Fujifilm) and normalized to the mean of the original non-normalized control values (HEK293 cells). A two-sided Student’s t-test was used to evaluate proteins statistically significantly regulated, with a p-value less than 0.05 that was set as the significance threshold.

Scilabra S.D., Pigoni M., Pravatá V., Schätzl T., Müller S.A., Troeberg L, & Lichtenthaler S.F. (2018). Increased TIMP-3 expression alters the cellular secretome through dual inhibition of the metalloprotease ADAM10 and ligand-binding of the LRP-1 receptor. Scientific Reports, 8, 14697.

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Quantifying Secreted Metabolites and Proteins from RPE Cells

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After incubating Transwells in 200 μL of RPE media both apically and basolaterally for 24 hours, 50 μL of supernatant from each side of the Transwell was analyzed using a commercial assay kit (Amplite Fluorimetric Beta-Hydroxybutyrate, #13831; AAT Bioquest, Sunnyvale, CA, USA) according to manufacturer's instructions. Standard curves were created using RPE media. Signal was read 20 minutes after mixing, using a luminometer (Promega).
ApoE, TIMP3, and PEDF secretion were measured from Transwells also incubated with 200 μL RPE media apically and basolaterally. Media was collected after a 72-hour incubation for ApoE and after a 24-hour incubation for TIMP3 and PEDF. A supernatant volume of 7.5 μL per well was subjected to PAGE and blotted with anti-ApoE (#AB947, Millipore Corp., Burlington, MA, USA), anti-TIMP3 (#AB6000, Millipore Corp.), or anti-PEDF (#AB180711; Abcam, Cambridge, UK).

Zhang Q., Presswalla F., Calton M., Charniga C., Stern J., Temple S., Vollrath D., Zacks D.N., Ali R.R., Thompson D.A, & Miller J.M. (2019). Highly Differentiated Human Fetal RPE Cultures Are Resistant to the Accumulation and Toxicity of Lipofuscin-Like Material. Investigative Ophthalmology & Visual Science, 60(10), 3468-3479.

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