IL-1 beta/IL-1F2 Overexpression Lysate

Name: IL-1 beta/IL-1F2 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-04165
Price: 689 USD
Availability: InStock

Novus Biologicals | Catalog # NBP2-04165

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for IL-1 beta/IL-1F2 Overexpression Lysate

IL-1 beta/IL-1F2 Transient Overexpression Lysate

Expression Host: HEK293T

Plasmid: RC202079

Accession#: NM_000576

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.

Theoretical MW

30.6 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 IL-1 beta/IL-1F2 Overexpression Lysate

Western Blot: IL-1 beta/IL-1F2 Overexpression Lysate [NBP2-04165]

Western Blot: IL1 beta Overexpression Lysate (Adult Normal) [NBP2-04165] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for IL1 beta.

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: IL-1 beta/IL-1F2

Interleukin-1 beta (IL-1 beta), also called IL-1F2, is a proinflammatory cytokine that functions in host defense following injury or inflammation (1-5). IL-1 beta is expressed by cells of the immune system such as monocytes, macrophage, neutrophils, and hepatocytes (1-3). Human IL-1 beta protein is expressed in the cytoplasm in an inactive precursor form (pro-IL-1 beta) which is 269 amino acids in length with a theoretical molecular weight of 31 kDa (1,4,6). Pro-IL-1 beta is cleaved by IL-1 converting enzyme (ICE), also called caspase-1, generating the 18 kDa mature, active protein that is secreted from cells (1,4,6). Pro-IL-1 beta is produced as a cellular response to molecular signals from pathogens and other processes termed pathogen associated molecular patterns (PAMPs) and damage associated molecular patterns (DAMPs) (1,2). PAMPs and DAMPs signal through pattern recognition receptors (PRRs) to induce formation of the inflammasome (1,2). The inflammasome is a multi-protein complex made of a PRR, such as Nod-like receptor family, pyrin domain containing 3(NLRP3), an adaptor, such as apoptosis-associated spec-like protein containing a CARD (ASC), and pro-caspase-1 (1-3,5). Assembly of the inflammasome enables cleavage of pro-caspase-1 into active caspase 1, followed by secretion of activate IL-1 beta (1-3,5).

IL-1 beta binding to its receptor IL-1RI and the downstream signaling contributes to a dual pathophysiological role (3). On one hand, IL-1 beta signaling activates immune cells and drives CD4+ T cell polarization to T helper type 1 (Th1) and Th17 cells, resulting in anti-tumor responses and mediation of acute inflammation (2,3). However, IL-1 beta also supports tumor growth and metastasis driven by multiple mechanisms including chronic inflammation, an immunosuppressive tumor microenvironment (TME), and angiogenesis (3). Additionally, IL-1 beta signaling been implicated in the pathogenesis of neuroinflammatory diseases of the central nervous system (CNS) such as multiple sclerosis (MS), Alzheimer's disease, and diabetic retinopathy (DR) (2). Mouse studies have shown regression of tumors treated with IL-1 as well as protective effects of IL-1 beta in instances of induced colitis and colon carcinoma (3). Conversely, blocking IL-1 beta has also shown promising effect in cancer treatment, especially when combined with chemotherapeutics (2,3). Approved IL-1 beta monoclonal antibody canakinumab has shown significant therapeutic promise in the treatment of DR (2). Given its multifaceted role in disease, IL-1 beta is a promising therapeutic target.

References

1. Lopez-Castejon G, Brough D. Understanding the mechanism of IL-1beta secretion. Cytokine Growth Factor Rev. 2011;22(4):189-195. https://doi.org/10.1016/j.cytogfr.2011.10.001

2. Mendiola AS, Cardona AE. The IL-1beta phenomena in neuroinflammatory diseases. J Neural Transm (Vienna). 2018;125(5):781-795. https://doi.org/10.1007/s00702-017-1732-9

3. Bent R, Moll L, Grabbe S, Bros M. Interleukin-1 Beta-A Friend or Foe in Malignancies?. Int J Mol Sci. 2018;19(8):2155. https://doi.org/doi:10.3390/ijms19082155

4. Krumm B, Xiang Y, Deng J. Structural biology of the IL-1 superfamily: key cytokines in the regulation of immune and inflammatory responses. Protein Sci. 2014;23(5):526-538. https://doi.org/10.1002/pro.2441

5. He Y, Hara H, Nunez G. Mechanism and Regulation of NLRP3 Inflammasome Activation. Trends Biochem Sci. 2016;41(12):1012-1021. https://doi.org/10.1016/j.tibs.2016.09.002

6. Uniprot (P01584)

Long Name

Interleukin 1 beta

Alternate Names

IL-1b, IL-1F2, IL1 beta, IL1B

Gene Symbol

IL1B

Additional IL-1 beta/IL-1F2 Products

Product Documents for IL-1 beta/IL-1F2 Overexpression Lysate

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Product Specific Notices for IL-1 beta/IL-1F2 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.