Hapln1

Manufactured by OriGene

Sourced in Germany

HAPLN1 is a protein-coding gene that produces the Hyaluronan and proteoglycan link protein 1. This protein is involved in the stabilization of proteoglycan aggregates in the extracellular matrix.

Automatically generated - may contain errors

Lab products found in correlation

Hrp conjugated secondary anti rabbit igg, by Santa Cruz Biotechnology (1 mentions) Imagequant las 4000, by GE Healthcare (1 mentions) Anti rabbit secondary antibody, by Jackson ImmunoResearch (1 mentions) Qubit assay kit, by Thermo Fisher Scientific (1 mentions) Las 4000, by Cytiva (1 mentions)

2 protocols using hapln1

Western Blot Analysis of HAPLN1 Secretion

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Briefly, supernatants of Lipofectamine 2000 alone, eGFP mRNA, or Link N mRNA-transfected SCP1 cells and chondrocytes were collected after 24 h. Protein concentrations were determined with the Lowry assay and 10 μg of cell supernatant was pipette onto the nitrocellulose membrane and allowed to air dry. Membranes were then blocked with 5% BSA in TBST for 1 h, followed by 3 washing steps (10 min each). Then, the membranes were incubated overnight with HAPLN1 (TA325115-Origene, Herford, Germany) primary antibody (dilution 1:1000) at 4 °C, and the next day membranes were washed 3 times (10 min each). Afterwards, the membrane was treated with HRP-conjugated secondary anti-rabbit IgG (Santa cruz, dilution 1:10,000) for 2 h. Membranes were developed upon incubating them with ECL substrate solution for a minute. Chemiluminescent signals were quantified using image J analysis software

Tendulkar G., Ehnert S., Sreekumar V., Chen T., Kaps H.P., Golombek S., Wendel H.P., Nüssler A.K, & Avci-Adali M. (2019). Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response. International Journal of Molecular Sciences, 20(7), 1716.

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Western Blot Analysis of Melanoma Samples

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Melanoma samples were harvested in appropriate amounts of whole cell lysis buffer (6% SDS, 30% glycerol, 0.18M Tris pH6.8, 5% DTT, bromophenol blue). Protein was quantified using Qubit assay kit (#Q33212, ThermoScientific) and 50μg protein was boiled for 10 minutes at 95°C and loaded on 4-20% Bis-Tris gels. Samples were run at 165V and transferred on PVDF membranes. Membranes were blocked for 1 hour in 5% milk and incubated with appropriate antibody overnight at 4°C. Next day, membranes were washed in TBS with 0.1% Tween and incubated with secondary antibody for 1 hour at room temperature. Membranes were washed and developed using chemiluminescence based detection reagents (#PI80196, ThermoScientific). Chemiluminescence was detected using ImageQuant^™ LAS 4000 (GE Healthcare Life Sciences, Pittsburgh, PA). Primary antibody to HSP90 was purchased from Cell Signaling (1:4000, 4877S), HAPLN1 (1:500, TA325115, Origene) and anti-rabbit secondary antibody was purchased from Jackson Immunoresearch (#211-032-171).

Kaur A., Ecker B.L., Douglass S.M., Kugel CH I.I.I., Webster M.R., Almeida F.V., Somasundaram R., Hayden J., Ban E., Ahmadzadeh H., Franco-Barraza J., Shah N., Mellis I.A., Keeney F., Kossenkov A., Tang H.Y., Yin X., Liu Q., Xu X., Fane M., Brafford P., Herlyn M., Speicher D.W., Wargo J.A., Tetzlaff M.T., Haydu L.E., Raj A., Shenoy V., Cukierman E, & Weeraratna A.T. (2018). Remodeling of the collagen matrix in aging skin promotes melanoma metastasis and affects immune cell motility. Cancer discovery, 9(1), 64-81.

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