Mabs1251

Manufactured by Merck Group

MABS1251 is a laboratory instrument designed for the analysis and characterization of monoclonal antibodies (mAbs). The device utilizes advanced analytical techniques to provide detailed information about the structure, purity, and stability of mAb samples.

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Lab products found in correlation

Mabn762, by Merck Group (2 mentions) Ph 9 buffer, by Vector Laboratories (1 mentions) Pathscan enabler 4, by Meyer Instruments (1 mentions) Envision flex hrp system, by Agilent Technologies (1 mentions)

5 protocols using mabs1251

Immunohistochemistry of Adrenal Enzymes

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IHC was performed on 10% FFPE tissue sections as described previously (21 (link)). The following primary antibodies were used: CYP11B2 (MilliporeSigma, MABS1251; diluted 1:1250; RRID, AB_2783793), 17α-hydroxylase/17, 20 lyase (CYP17A1) (LSBio, LS-B14227; diluted 1:2000; RRID, AB_2857939), 11β-hydroxylase (CYP11B1) (clone 80-7-3; kindly provided by Dr. Celso Gomez-Sanchez; diluted 1:50; RRID, AB_2650563), and visinin like 1 (VSNL1) (MilliporeSigma, MABN762; diluted 1:1000; RRID, AB_2832208).

Nanba K., Blinder A.R., Udager A.M., Hirokawa Y., Miura T., Okuno H., Moriyoshi K., Yamazaki Y., Sasano H., Yasoda A., Satoh-Asahara N., Rainey W.E, & Tagami T. (2024). Double somatic mutations in CTNNB1 and GNA11 in an aldosterone-producing adenoma. Frontiers in Endocrinology, 15, 1286297.

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Immunohistochemical Analysis of Adrenal Gland Tumor Markers

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APA FFPE sections were deparaffinized and epitope retrieval was performed by heating samples for 15 minutes in a pH 9 buffer (Vector Laboratories Inc). CYP11B2 and 17α-hydroxylase/17,20 lyase (CYP17A1) IHC on the APAs was performed as previously described [14 (link)]. After peroxidase blocking, APA FFPE sections were incubated overnight at 4^oC with antihuman mouse monoclonal antibodies against CYP11B2 (clone 41-17B; diluted 1:100; Millipore Sigma; catalog No. MABS1251) [15 ] and visinin-like 1 (VSNL1) (diluted 1:1000; Millipore Sigma; catalog No. MABN762) [16 ], a rat monoclonal antibody against human 11β-hydroxylase (CYP11B1) (diluted 1:100; from Dr Gomez-Sanchez) [17 ]; and at room temperature for 1 hour with an antihuman rabbit polyclonal antibody against CYP17A1 (diluted 1:2000; LifeSpan Biosciences, catalog No. LS-B14227) [18 ]. The Polink-2 HRP Plus Mouse DAB System (GBI Labs) was used for detection. Slides were counterstained with Harris hematoxylin for 10 to 20 seconds followed by dehydration and cover-slipping.

Rege J., Nanba K., Blinder A.R., Plaska S., Udager A.M., Vats P., Kumar-Sinha C., Giordano T.J., Rainey W.E, & Else T. (2020). Identification of Somatic Mutations in CLCN2 in Aldosterone-Producing Adenomas. Journal of the Endocrine Society, 4(10), bvaa123.

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Immunohistochemistry of Adrenal Glands

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Tissue samples of 8 adrenal glands from the 5 included patients evaluated and treated at our institution were analyzed with immunohistochemistry. Formalin-fixed, paraffin-embedded tissue sections were cut to a thickness of 5 µm. Immunohistochemistry was performed using primary antibodies against aldosterone synthase (CYP11B2) and tyrosine hydroxylase as markers for aldosterone and catecholamine producing cells, respectively. The antibodies used in immunohistochemistry were as follows: mouse monoclonal antibody against CYP11B2 (MABS1251 from Millipore/Sigma, 1:1250) [20 (link)] and a rabbit polyclonal antibody against tyrosine hydroxylase (Provided by Dr. John Porter, 1:15 000) [21 (link)]. The reacted slides were counterstained with Harris hematoxylin and then dehydrated and coverslipped.

Mao J.J., Baker J.E., Rainey W.E., Young WF J.r, & Bancos I. (2021). Concomitant Pheochromocytoma and Primary Aldosteronism: A Case Series and Literature Review. Journal of the Endocrine Society, 5(8), bvab107.

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Immunohistochemical Screening of Adrenal Zones

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Antibodies toward steroid or catecholamine synthetic enzymes with adrenal zone-selective expression (Figure 1) were used for IHC to screen candidate adrenals for zone capture. IHC staining was performed using antibodies against CYP11B2 (mouse monoclonal clone 41–17B; 1:1250; Millipore; number MABS1251) (Gomez-Sanchez et al., 2014 (link)), VSNL1 (mouse monoclonal; 1:1000; Millipore; number MABN762), HSD3B2 (mouse monoclonal; 1:5000, kindly provided by Dr. Gomez-Sanchez) (Gomez-Sanchez et al., 2017 (link)), CYB5A (mouse monoclonal; 1:5000; Acris; number AM31963PU-N), and TH (rabbit monoclonal; 1:15,000, kindly provided by Dr. John Porter) (Porter, 1986 (link)) for 60 minutes on FFPE sections. Tissue sections were boiled for 15 min in Tris/EDTA pH 9 antigen retrieval buffer (Vector Laboratories, Burlingame, CA). The FLEX HRP EnVision System (Dako) was used for detection. Diaminobenzidine chromogen was then applied for approximately 45 seconds, 30 seconds, 30 seconds, 30 seconds, and 15 seconds respectively. Slides were counterstained with Harris hematoxylin for 5 seconds and then dehydrated and coverslipped. IHC images were scanned using PathScan Enabler IV (Meyer Instruments, Houston, TX).

Baker J.E., Plaska S.W., Qin Z., Liu C.J., Rege J., Rainey W.E, & Udager A.M. (2021). Targeted RNA sequencing of adrenal zones using immunohistochemistry-guided capture of formalin-fixed paraffin-embedded tissue. Molecular and cellular endocrinology, 530, 111296.

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Immunohistochemistry of Steroidogenic Enzymes

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Formalin-fixed paraffin-embedded tissue blocks were available from all but one patient (Case 10, APA-Y182). Immunohistochemistry (IHC) was performed using specific antibodies against CYP11B2 (mouse monoclonal, Millipore, #MABS1251) (Gomez-Sanchez, et al. 2014 (link)), 11β-hydroxylase (CYP11B1, rat monoclonal) (Gomez-Sanchez, et al. 2017 (link)), 17α-hydroxylase (CYP17A1, mouse monoclonal) (Dharia, et al. 2004 (link)), 3β-hydroxysteroiddehydrogenase/Δ4–5 isomerase type II (HSD3B2, mouse monoclonal) (Gomez-Sanchez et al. 2017 (link)), and β-catenin (mouse monoclonal, BD Biosciences, #610153) essentially as described previously (Monticone, et al. 2012 (link); Nanba et al. 2016 (link)).

Lerario A.M., Nanba K., Blinder A.R., Suematsu S., Omura M., Nishikawa T., Giordano T.J., Rainey W.E, & Else T. (2019). Genetics of aldosterone-producing adenomas with pathogenic KCNJ5 variants. Endocrine-related cancer, 26(4), 463-470.

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