MCPIP1/ZC3H12A Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-06242

Novus Biologicals, part of Bio-Techne
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NBP2-06242
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for MCPIP1/ZC3H12A Overexpression Lysate

MCPIP1/ZC3H12A Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC201381

Accession#: NM_025079

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

65.5 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for MCPIP1/ZC3H12A Overexpression Lysate

Western Blot: MCPIP1/ZC3H12A Overexpression Lysate [NBP2-06242]

Western Blot: MCPIP1/ZC3H12A Overexpression Lysate [NBP2-06242]

Western Blot: ZC3H12A Overexpression Lysate (Adult Normal) [NBP2-06242] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for ZC3H12A.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: MCPIP1

Monocyte chemoattractant protein-induced protein 1 (MCPIP1), also referred to as ZC3H12A and Regnase-1, is an endonuclease that cleaves and destabilizes mRNA and functions as a negative regulator of inflammation (1,2). MCPIP1 is encoded by the ZC3H12A gene as a protein of 599 amino acids (aa) in length with a theoretical molecular weight of approximately 66 kDa (3-5). Common structural features of the MCPIP1 protein are the ubiquitin-associated (UBA) domain (43-89 aa), two protein-rich regions (PRRs) (100-126 aa, 458-536 aa), the PilT N-terminal (PIN) domain (133-270 aa), the zinc-finger motif (305-325 aa), the natively disordered region (326-457 aa), and the C-terminal conserved region (CCR) (545-598 aa) (1,3-5). MCPIP1 functions as an anti-inflammatory protein with RNase activity that is known to degrade many cytokine mRNA transcripts including interleukin-1beta (IL-1beta), IL-6, and IL-17 (4,5). In addition to its regulation of inflammation through mRNA degradation, MCPIP1 also controls inflammation through ubiquitin-specific peptidase 10 (USP10)-mediated protein deubiquitination of TRAF6 which results in suppression of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NFkappaB) transcription factors (1,2,5). Besides regulating inflammation, MCPIP1 is involved in several non-immune cellular processes including apoptosis, adipogenesis, angiogenesis, and proliferation (2,5). MCPIP1 has also been implicated in a number of autoimmune diseases and other pathologies including chronic obstructive pulmonary disease (COPD), diabetes, rheumatoid arthritis, and cancer (3,5). Additionally, MCPIP1/ZC3H12A has been shown to have anti-viral activity against RNA viruses such as Japanese encephalitis virus and human immunodeficiency virus (HIV)-1 (1,5).

References

1. Uehata, T., & Takeuchi, O. (2017). Regnase-1 Is an Endoribonuclease Essential for the Maintenance of Immune Homeostasis. Journal of interferon & cytokine research: the official journal of the International Society for Interferon and Cytokine Research, 37(5), 220-229. https://doi.org/10.1089/jir.2017.0001

2. Miekus, K., Kotlinowski, J., Lichawska-Cieslar, A., Rys, J., & Jura, J. (2019). Activity of MCPIP1 RNase in tumor associated processes. Journal of experimental & clinical cancer research : CR, 38(1), 421. https://doi.org/10.1186/s13046-019-1430-6

3. Jura, J., Skalniak, L., & Koj, A. (2012). Monocyte chemotactic protein-1-induced protein-1 (MCPIP1) is a novel multifunctional modulator of inflammatory reactions. Biochimica et biophysica acta, 1823(10), 1905-1913. https://doi.org/10.1016/j.bbamcr.2012.06.029

4. Xu, J., Fu, S., Peng, W., & Rao, Z. (2012). MCP-1-induced protein-1, an immune regulator. Protein & cell, 3(12), 903-910. https://doi.org/10.1007/s13238-012-2075-9

5. Musson, R., Szukala, W., & Jura, J. (2020). MCPIP1 RNase and Its Multifaceted Role. International journal of molecular sciences, 21(19), 7183. https://doi.org/10.3390/ijms21197183

Long Name

MCP-1-induced Protein 1

Alternate Names

ZC3H12A

Gene Symbol

ZC3H12A

Additional MCPIP1 Products

Product Documents for MCPIP1/ZC3H12A Overexpression Lysate

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for MCPIP1/ZC3H12A Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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