Q: Can I distribute the solution as 10 ul per tube, 10 tubes in total, and store them in -80 degree Celsius refrigerator?

A: Yes, that should be okay, although we tend to recommend storing this antibody at -20 degrees Celsius. Aliquoting is a good idea because freeze-thaw cycles should be avoided.

Q: Do you have a protocol for guiding me how to prepare the solution of this antibody?

A: Our reconstitution instructions for the lyophilized antibody NBP1-42767 (IL1 beta Antibody) are 'restore with 100 ul deionized water (or equivalent)'.

IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

Novus Biologicals | Catalog # NBP1-42767

(1)

Citations (13)

Product Documents

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Product Specifications
Scientific Data
Applications
Flow Cytometry
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Species Reactivity

Validated:

Mouse, Rat

Cited:

Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, ELISA, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Dot Blot, CyTOF-ready

Cited:

Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

Azide and BSA Free

View all IL-1 beta/IL-1F2 Primary Antibodies »

Product Specifications

Immunogen

This IL-1 beta/IL-1F2 Antibody was prepared by repeated immunizations with recombinant mouse IL-1 beta/IL-1F2 produced in E.coli. The MW of recombinant mouse IL-1 beta/IL-1F2 was 17 kDa. (Uniprot: P10749)

Specificity

This antibody is primarily directed against mature, 17,000 MW mouse IL-1beta and is useful in determining its presence in various assays. The antibody does not recognize human IL-1beta or mouse IL-1alpha based on a neutralization assay. In ELISA formats and other immunoreactive assays, reactivity occurs with rat IL-1beta. This antibody will recognize 10% of the non-denatured (native) precursor 31,000 MW mouse IL-1beta containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in immunoblot analysis, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1beta precursor molecule. Denatured 31,000 precursor IL-1beta will be recognized by this antibody.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

This is an IgG preparation of whole rabbit serum purified by DEAE fractionation

Store this antibody at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767]

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Analysis using the HRP conjugate of NBP1-42767. Detection of Lane 1: Mouse IL-1 Beta Load: 50 ug per Detection of Lane Secondary antibody: IL1 beta Antibody Peroxidase conjugated at 1:1,000 for 30 min at RT Block for30 min RT Predicted/Observed size: 18 kDa, 18 kDa.

Immunocytochemistry/ Immunofluorescence: IL-1 beta/IL-1F2 Antibody [NBP1-42767]

Immunocytochemistry/Immunofluorescence: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Staining of mouse carotid artery. A) No Ab staining of WT uninjured Ms. B) IL-1 beta staining of cells after surgical injury of tissue. C) No Ab staining of injured tissue from an IL-1 beta KO.

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody [NBP1-42767]

Immunohistochemistry: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Analysis of Mouse Embryonic Kidney Tissue: Mouse Embryonic Kidney Fixation: FFPE buffered formalin 10% conc Ag Retrieval: Heat, Citrate pH 6.2. Pressure Cooker Primary antibody: 2ug/ml 1.5 hour @ room T Secondary Ab: MACH 1 HRP POLYMER 1:50 45C RT

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767]

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Analysis using the Biotin conjugate of NBP1-42767. Detection of Lane 1: Mouse IL-1beta Recombinant Protein. Load: 50 ng per lane. Used at a dilution of 1:1,000 overnight at 4C.

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767]

Western Blot: IL-1 beta/IL-1F2 Antibody [NBP1-42767] - Detects 10% of the non-denatured (native) precursor 31,000 MW mouse IL-1Beta containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in western blot analysis, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1Beta precursor molecule. Denatured IL-1Beta will have a 18 kDa band.

IL-1 beta/IL-1F2 Antibody

Immunohistochemistry of Rabbit anti-IL-1 beta/IL-1F2 Antibody in Mouse Embryonic Kidney Tissue: Mouse Embryonic Kidney Fixation: FFPE buffered formalin 10% conc Ag Retrieval: Heat, Citrate pH 6.2. Pressure Cooker Primary antibody: 2ug/ml 1.5 hour @ room T Secondary Ab: MACH 1 HRP POLYMER 1:50 45" RT

IL-1 beta/IL-1F2 Antibody

This antibody will recognize 10% of the non-denatured (native) precursor 31,000 MW mouse IL-1beta containing samples but will primarily detect all of the 17,000 MW mature molecule. However, in western blot analysis, the usual procedure of heating the sample in SDS with or without reducing agents will facilitate denaturing of the 31,000 MW IL- 1beta precursor molecule. Denatured IL-1beta will have a 18 kDa band.

Western Blot: IL-1 beta/IL-1F2 Antibody - Azide and BSA Free [NBP1-42767] -

HIV Vpu and gp41 play a role in inflammasome activation.Monocyte derived macrophages (MDMs) (1 x 106) were either left untreated or transfected with 2μg of pVpu plasmid expressing Vpu or pgp41 or gp120 plasmid expressing gp120. Supernatants were collected and tested for IL-1 beta and IL-18 secretion at 12 hr (A) and 24 hr (B). Cells extracts at 12 hr from mock or transfected cells were analysed for the presence of caspase-1 p20 by western blotting, followed by quantification using Image Studio Lite (Licor) and normalized to internal control ( beta -actin) from 3 western blots(C). MDMs expressing gp41 (D) or Vpu (E) monocytes were silenced for TLR2, TLR3, TLR4 and TLR8 by siRNA. Supernatant was collected at 12 hr and analysed for IL-1 beta and IL-18 using ELISA. Cell extracts of MDMs stimulated with either Vpu (F) or gp41 (G) were analysed for the presence of pro-IL-1 beta, pro-IL-18, cleaved IL-1 beta or cleave IL-18 by western blotting, followed by quantification using Image Studio Lite (Licor) and normalized to internal control ( beta -actin) from 3 western blots. MDMs expressing Vpu (H-J), gp41 (K-M) or pgmock (N-P) were silenced for NLRP3, NLRP1, NLRC4 or NLRP3 and NLRC4 by siRNA. Supernatant was collected at 12 hr and analysed for IL-1 beta (Η, Κ, Ν) and IL-18 (I,L,O) using ELISA. All graphs were normalized to the level of siRNA knockdown. Cells extracts were analysed for the presence of caspase-1 p20 at 12 hr by western blotting, followed by quantification using Image Studio Lite (Licor) and normalized to internal control ( beta -actin) from 3 western blots (J,M,P). The data represent the mean of three independent experiments +/- SD (n = 3 sets of macrophages) yielding consistent results. **, p < 0.005 and ***, p < 0.001 indicate statistically significant differences. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33861800), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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Applications for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

Application

Recommended Usage

ELISA

1:1000-1:5000

Flow Cytometry

1:10-1:1000

Immunocytochemistry/ Immunofluorescence

1:50-1:250

Immunohistochemistry

1:50-1:250

Immunohistochemistry-Paraffin

1:10-1:500

Immunoprecipitation

1:200-1:800

Western Blot

1:500-1:2000

Application Notes

This product has been tested for use in immunohistochemistry, immunoblotting and immunofluorescence. This antibody is useful in ELISA, neutralizations, radioimmunoassays, flow cytometry, and immunoprecipitation. It recognizes the 17,000 MW mature IL-1beta. For immunoblots, typically, IL-1beta is detected from supernatants or lysates of 2 x 10E6 endotoxin-stimulated peripheral blood mononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1% (v/v) serum plus 10 ng/mL E.coli LPS. For immunoprecipitation pre-clearing the preparation with a non-specific Rabbit IgG to reduce background is suggested. For immunohistochemistry either paraffin fixation or cryofixation can be used for sample preparation to stain intracellular IL-1beta. For ELISA use HRP Conjugated Anti-Rabbit IgG [H&L] (Goat) (611-1302) for detection. In ELISA formats this antibody is best used as the second antibody in combination with a monoclonal antibody as a capture antibody. This antibody is also useful for neutralization of mouse and rat IL-1beta activity in bioassays. It does not neutralize the biological activity IL-1alpha. It does not neutralize the biological activity of human or primate IL-1beta. For neutralization, it is recommended to incubate the sample with a dilution of the antibody for at least 4 hours before being tested. A control of similarly diluted normal rabbit IgG is recommended. This antibody can be used for FACS analysis. Caution should be exhibited as the F(c) domain of the rabbit IgG molecule may interact with cells non-specifically.

Reviewed Applications

Read 1 review rated 5 using NBP1-42767 in the following applications:

Flow Cytometry (1 Review)

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Formulation, Preparation, and Storage

Purification

Column Chromatography

Reconstitution

Reconstitute with 100 ul deionized water (or equivalent)

Formulation

Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Format

Azide and BSA Free

Preservative

No Preservative

Concentration

LYOPH mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store lyophilized antibody at 4C. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

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

Entrez Gene IDs

16176 (Mouse); 24494 (Rat)

Gene Symbol

IL1B

UniProt

P10749

Additional IL-1 beta/IL-1F2 Products

Product Documents for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

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Certificate of Analysis

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Product Specific Notices for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Cancer Biomarkers

Conventional/Classical Dendritic Cells

Cytokines and Receptors in Allergy and Asthma

Cytokines and Receptors in Angiogenesis

Group 3 ILCs

IL-1 Family

Infectious Diseases

Inflammatory Mediators

Isolation, Culture, and Identification of Natural Killer Cells

LTi Cells

Macrophage Activation Markers

Microglia Activation Markers

Myeloid-derived Suppressor Cells (MDSC)

NOD-like Receptors and the Inflammasome

Regulatory T Cells (Tregs)

T Cell Cytokine Signaling

Citations for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

Customer Reviews for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free (1)

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IL-1 beta/IL-1F2 Antibody

Name: James Xiao

Application: Flow Cytometry

Sample Tested: Lung tissue

Species: Mouse

Verified Customer | Posted 01/23/2017

Left: IL-1b-stained sample; left panel: isotype control.

Single cells were isolated from a mouse lung graft and stained with IL-1b ab (1:00 in FACS buffer) for 30 minutes. Cells were then washed and stained with Goat anti-rabbit FITC-conjugated secondary antibody for additional 30 minutes. Cells were then washed and analyzed by flow cytometry.

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Protocols

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FAQs for IL-1 beta/IL-1F2 Antibody - Azide and BSA Free

Showing 1 - 2 of 2 FAQs Showing All

Q: Can I distribute the solution as 10 ul per tube, 10 tubes in total, and store them in -80 degree Celsius refrigerator?

A: Yes, that should be okay, although we tend to recommend storing this antibody at -20 degrees Celsius. Aliquoting is a good idea because freeze-thaw cycles should be avoided.
Q: Do you have a protocol for guiding me how to prepare the solution of this antibody?

A: Our reconstitution instructions for the lyophilized antibody NBP1-42767 (IL1 beta Antibody) are 'restore with 100 ul deionized water (or equivalent)'.
Q: Can I distribute the solution as 10 ul per tube, 10 tubes in total, and store them in -80 degree Celsius refrigerator?

A: Yes, that should be okay, although we tend to recommend storing this antibody at -20 degrees Celsius. Aliquoting is a good idea because freeze-thaw cycles should be avoided.
Q: Do you have a protocol for guiding me how to prepare the solution of this antibody?

A: Our reconstitution instructions for the lyophilized antibody NBP1-42767 (IL1 beta Antibody) are 'restore with 100 ul deionized water (or equivalent)'.