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H-129 is a laboratory instrument designed for the detection and analysis of specific proteins or molecules in biological samples. It utilizes advanced spectroscopic techniques to provide accurate and reliable results. The core function of H-129 is to facilitate the identification and quantification of target analytes, enabling researchers to obtain crucial data for their scientific investigations.

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Lab products found in correlation

Magna chip g chromatin immunoprecipitation kit, by Merck Group (2 mentions) Hmec cells, by Lonza (2 mentions) β actin, by Merck Group (1 mentions) Ab23980, by Abcam (1 mentions) H3k27ac, by Diagenode (1 mentions) C15410174, by Diagenode (1 mentions) β tubulin, by Merck Group (1 mentions) H 137, by Santa Cruz Biotechnology (1 mentions) Anti myc 9b11, by Cell Signaling Technology (1 mentions) Complete protease inhibitor cocktail, by Roche (1 mentions) Kapa hyper prep kit, by Roche (1 mentions) Nextseq 500, by Illumina (1 mentions)

Close spelling variants of this product

GRP78 (H-129, Anti-GRP-78 H-129 Primary anti-LNK (H-129,

7 protocols using h 129

1

Protein Expression Analysis by Western Blot

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Total cell lysates were separated by SDS-PAGE and analyzed by western blotting using antibodies against p63 (Santa Cruz; H-129), NRF2 (Santa Cruz; C-20), K10 (DAKO) and β-actin (Sigma).
Kurinna S., Seltmann K., Bachmann A.L., Schwendimann A., Thiagarajan L., Hennig P., Beer H.D., Mollo M.R., Missero C, & Werner S. (2021). Interaction of the NRF2 and p63 transcription factors promotes keratinocyte proliferation in the epidermis. Nucleic Acids Research, 49(7), 3748-3763.
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2

ChIP-qPCR analysis of FZD7 regulation

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HMEC cells (Lonza) or MMTV-Wnt-1 primary cells were grown to 80% confluence and cells were then cross-linked with 1% formaledyde and processed. The crosslinking, immunoprecipitation, washing, elution, reverse crosslinking, and proteinase K treatment were performed according to the manufacturer’s directions described in the Magna ChIP G Chromatin Immunoprecipitation Kit from Millipore. Antibodies used were anti-4A4, H-129 (Santa Cruz) or normal rabbit IgG. Purified immunoprecipitated DNA was used for real time qPCR. Primers for ChIP PCR are: 5′-TATCAGCATTCCAGGCCCAC-3′ (forward) and 5′-TTCCTGGGAGAACAATCGCC-3′ (reverse) for human FZD7; 5′-AATGAGGCAAACACCCCCTC-3′ (forward) and 5′-CTCCGGGGGATTAAAGGTGG-3′ (reverse) for mouse Fzd7; 5′-CGAGATCCCTCCAAAATCAA (forward) and 5′-CCCAGCCTTCTCCATGG (reverse) for human GAPDH and 5′-AACATCAAATGGGGTGAGG-3′ (forward), 5′-GGCCTTCTCCATGGTGGT-3′ (reverse) for mouse Gapdh.
Chakrabarti R., Wei Y., Hwang J., Hang X., Blanco M.A., Choudhury A., Tiede B., Romano R.A., DeCoste C., Mercatali L., Ibrahim T., Amadori D., Kannan N., Eaves C.J., Sinha S, & Kang Y. (2014). ΔNp63 promotes stem cell activity in mammary gland development and basal-like breast cancer by enhancing Fzd7 expression and Wnt signaling. Nature cell biology, 16(10), 1004-13.
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3

Profiling Epidermal Differentiation via ChIP-seq

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Human primary keratinocytes under proliferating, early, mid-, and late differentiation condition generated from HKC1 were used for ChIP and ChIP-seq analysis. ChIP-sequencing datasets were generated with an antibody recognizing the alpha isoforms of p63 (H129, Santa Cruz), an antibody that detects all forms of the large subunit of RNA polymerase II (8WG16, Santa Cruz), and a H3K27ac antibody recognizing the acetylation of the lysine 27 residue of histone H3 (H3K27ac, C15410174, pAb-174-050, Diagenode). ChIP-qPCR experiments were performed using 2 μg RUNX1 antibody (ab23980, Abcam) and 1 μg TFAP2 antibody (C18X, Santa cruz). RNAPII, p63, H3K27ac, RUNX1, and TFAP2 ChIP experiments were performed as previously described 65 (link), with a minor change of using magnetic beads (Novex by Life Technologies, 10008D and 10009D).
Kouwenhoven E.N., Oti M., Niehues H., van Heeringen S.J., Schalkwijk J., Stunnenberg H.G., van Bokhoven H, & Zhou H. (2015). Transcription factor p63 bookmarks and regulates dynamic enhancers during epidermal differentiation. EMBO Reports, 16(7), 863-878.
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4

Western Blot Analysis of p63 Isoforms

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Western blot analysis was performed as previously described [77 (link)]. Primary antibodies were used at the following dilutions: Pan-p63 (Nekulova et al. [54 (link)], 1:5000), H-129 (Santa Cruz, 1:5000), TAp63 (Nekulova et al. [54 (link)], 1:5000), ΔNp63 (Nekulova et al. [54 (link)], 1:5000) and beta-Tubulin (Sigma, 1:10,000).
Sethi I., Romano R.A., Gluck C., Smalley K., Vojtesek B., Buck M.J, & Sinha S. (2015). A global analysis of the complex landscape of isoforms and regulatory networks of p63 in human cells and tissues. BMC Genomics, 16, 584.
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5

Protein Detection in Zebrafish Cells

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Zebrafish embryos were dechorionated and deyolked, and cells were collected as described [84] (link). Cell pellets or adult tissues were either directly dissolved in SDS loading buffer as described [84] (link), or first lysed in chilled CSH buffer (50 mM Tris-HCl (pH 7.5), 250 mM NaCl, 1 mM EDTA, 1% Triton-X100, supplemented with cOmplete Protease Inhibitor Cocktail, Roche), followed by protein concentration determination. 10–12% SDS-PAGE, blotting on nitrocellulose membrane, Ponceau staining and immunodetection were carried out as described [84] (link). Used primary antibodies were: anti-Myc, 9B11 (mouse, Cell Signaling Technology; 1∶2000); anti-p63, 4A4 (mouse, Santa Cruz Technologies, against aa 1–205 of human ΔNp63), D-9 (mouse, Santa Cruz Biotechnology, against aa 15–151 of human ΔNp63), H-137 (rabbit, Santa Cruz Biotechnology, against aa 15–151 of human ΔNp63), H-129 (rabbit, Santa Cruz Biotechnology, against aa 513–641 at C-terminus of human TAp63α).
Fischer B., Metzger M., Richardson R., Knyphausen P., Ramezani T., Franzen R., Schmelzer E., Bloch W., Carney T.J, & Hammerschmidt M. (2014). p53 and TAp63 Promote Keratinocyte Proliferation and Differentiation in Breeding Tubercles of the Zebrafish. PLoS Genetics, 10(1), e1004048.
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6

ChIP-seq Profiling of Histone Marks

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Chromatin for ChIP was prepared as previously described [96 (link)] with minor modifications. On average, 0.5 M cells were used in each ChIP. Antibodies against H3K27ac (Diagenode #C15410174, 1.2 μg), H3K4me3 (Diagenode #C15410003, 1 μg), H3K27me3 (Diagenode #C15410069, 1.5 μg), p63 (Santa Cruz #H129, 1 μg, recognizing the C-terminal α tail of p63) were used in ChIP assay. Afterwards, 5 ng DNA fragments were pooled and proceeded on with library construction using KAPA Hyper Prep Kit (Kapa Biosystems #KK8504) according to the standard protocol. The prepared libraries were then sequenced using the NextSeq 500 (Illumina) according to standard Illumina protocols.
Smits J.G., Cunha D.L., Amini M., Bertolin M., Laberthonnière C., Qu J., Owen N., Latta L., Seitz B., Roux L.N., Stachon T., Ferrari S., Moosajee M., Aberdam D., Szentmary N., van Heeringen S.J, & Zhou H. (2023). Identification of the regulatory circuit governing corneal epithelial fate determination and disease. PLOS Biology, 21(10), e3002336.
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7

ChIP-qPCR analysis of FZD7 regulation

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HMEC cells (Lonza) or MMTV-Wnt-1 primary cells were grown to 80% confluence and cells were then cross-linked with 1% formaledyde and processed. The crosslinking, immunoprecipitation, washing, elution, reverse crosslinking, and proteinase K treatment were performed according to the manufacturer’s directions described in the Magna ChIP G Chromatin Immunoprecipitation Kit from Millipore. Antibodies used were anti-4A4, H-129 (Santa Cruz) or normal rabbit IgG. Purified immunoprecipitated DNA was used for real time qPCR. Primers for ChIP PCR are: 5′-TATCAGCATTCCAGGCCCAC-3′ (forward) and 5′-TTCCTGGGAGAACAATCGCC-3′ (reverse) for human FZD7; 5′-AATGAGGCAAACACCCCCTC-3′ (forward) and 5′-CTCCGGGGGATTAAAGGTGG-3′ (reverse) for mouse Fzd7; 5′-CGAGATCCCTCCAAAATCAA (forward) and 5′-CCCAGCCTTCTCCATGG (reverse) for human GAPDH and 5′-AACATCAAATGGGGTGAGG-3′ (forward), 5′-GGCCTTCTCCATGGTGGT-3′ (reverse) for mouse Gapdh.
Chakrabarti R., Wei Y., Hwang J., Hang X., Blanco M.A., Choudhury A., Tiede B., Romano R.A., DeCoste C., Mercatali L., Ibrahim T., Amadori D., Kannan N., Eaves C.J., Sinha S, & Kang Y. (2014). ΔNp63 promotes stem cell activity in mammary gland development and basal-like breast cancer by enhancing Fzd7 expression and Wnt signaling. Nature cell biology, 16(10), 1004-13.
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