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Mouse IL‑1 beta/IL‑1F2 Antibody

Catalog # AF-401-NA | R&D Systems, Inc. a Bio-Techne Brand
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AF-401-NA
AF-401-SP

Key Product Details

Species Reactivity

Mouse

Applications

Immunocytochemistry, Immunohistochemistry, Neutralization, Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Summary for Mouse IL‑1 beta/IL‑1F2 Antibody

Immunogen

E. coli-derived recombinant mouse IL-1 beta/IL-1F2
Val118-Ser269
Accession # NP_032387

Specificity

Detects mouse IL-1 beta/IL-1F2 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse IL‑1 beta/IL‑1F2 Antibody

Detection of Human and Mouse IL-1 beta/IL-1F2 antibody by Western Blot.

Detection of Human and Mouse IL‑1 beta/IL‑1F2 by Western Blot.

Western blot shows lysates of THP-1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS for 4 hours and RAW 264.7 mouse monocyte/macrophage cell line untreated (-) or treated (+) with 10 µg/mL LPS for 24 hours. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse IL-1 beta/IL-1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for IL-1 beta/IL-1F2 at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
IL-1 beta/IL-1F2 antibody in MCF-7 Human Cell Line by Immunocytochemistry (ICC).

IL‑1 beta/IL‑1F2 in MCF‑7 Human Cell Line.

IL-1 beta/IL-1F2 was detected in immersion fixed MCF-7 human breast cancer cell line using Goat Anti-Mouse IL-1 beta/IL-1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
IL-1 beta/IL-1F2 antibody in Mouse Thymus by Immunohistochemistry (IHC-Fr).

IL‑1 beta/IL‑1F2 in Mouse Thymus.

IL-1 beta/IL-1F2 was detected in perfusion fixed frozen sections of mouse thymus using Goat Anti-Mouse IL-1 beta/IL-1F2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-401-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Applications for Mouse IL‑1 beta/IL‑1F2 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed mouse T cells and immersion fixed MCF-7 human breast cancer cell line

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen Mouse Thymus and perfusion fixed paraffin-embedded sections of Mouse Intestine.

Simple Western

2.5 µg/mL
Sample: RAW 264.7 mouse monocyte/macrophage cell line treated with LPS

Western Blot

0.25 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line treated with PMA and LPS and RAW 264.7 mouse monocyte/macrophage cell line treated with LPS

Neutralization

Measured by its ability to neutralize IL‑1 beta/IL‑1F2-induced proliferation in the D10.G4.1 mouse helper T cell line. Symons, J.A. et al. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 272. The Neutralization Dose (ND50) is typically ≤0.25 µg/mL in the presence of 50 pg/mL Recombinant Mouse IL‑1 beta/IL‑1F2.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 24 reviews rated 4.1 using AF-401-NA in the following applications:

Formulation, Preparation and Storage

Purification

Antigen Affinity-purified

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

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Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-1 beta/IL-1F2

IL-1 is a name that designates two pleiotropic cytokines, IL-1 alpha (IL-1F1) and IL-1 beta (IL-1F2, IL1B), which are the products of distinct genes. IL-1 alpha and IL-1 beta are structurally related polypeptides that share approximately 17% amino acid (aa) identity in mouse. Both proteins are produced by a wide variety of cells in response to inflammatory agents, infections, or microbial endotoxins. While IL-1 alpha and IL-1 beta are regulated independently, they bind to the same receptor and exert identical biological effects. IL-1 RI binds directly to IL-1 alpha or IL-1 beta and then associates with IL-1 R accessory protein (IL-1 R3/IL-1 R AcP) to form a high-affinity receptor complex that is competent for signal transduction. IL-1 RII has high affinity for IL-1 beta but functions as a decoy receptor and negative regulator of IL-1 beta activity. IL-1ra functions as a competitive antagonist by preventing IL-1 alpha and IL-1 beta from interacting with IL-1 RI. Intracellular cleavage of the IL-1 beta precursor by Caspase-1/ICE is a key step in the inflammatory response. The 17 kDa molecular weight mature mouse IL-1 beta shares 90% aa sequence identity with cotton rat and rat and 67%-78% with canine, equine, feline, human, porcine, and rhesus macaque IL-1 beta. IL-1 beta functions in a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. IL-1 beta dysregulation is implicated in many pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin-dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases.

Long Name

Interleukin 1 beta

Alternate Names

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

Entrez Gene IDs

403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig); 3553 (Human); 16176 (Mouse); 397122 (Porcine); 24494 (Rat)

Gene Symbol

IL1B

UniProt

Product Documents for Mouse IL‑1 beta/IL‑1F2 Antibody

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse IL‑1 beta/IL‑1F2 Antibody

For research use only

Citations for Mouse IL‑1 beta/IL‑1F2 Antibody (269)

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Customer Reviews for Mouse IL‑1 beta/IL‑1F2 Antibody (24)

4.1 out of 5
24 customer ratings
5 stars
29%
4 stars
58%
3 stars
8%
2 stars
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1 star
4%

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Showing  1 - 5 of 24 reviews Showing All
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  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Aorta tissue
    Species: Mouse
    Verified Customer | Posted 07/25/2023
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Katie Myers
    Application: ELISA
    Sample Tested: Serum
    Species: Mouse
    Verified Customer | Posted 11/29/2022
    Used as ELISA detection
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Lysates of primary bone marrow derived macrophage
    Species: Mouse
    Verified Customer | Posted 11/08/2022
    IL-1beta band comes at 32 KDa for the pro-IL-1b.
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Leukocyte
    Species: Mouse
    Verified Customer | Posted 08/28/2022
    Works well with flow cytometry!
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Peripheral blood mononuclear cells (PBMCs)
    Species: Human
    Verified Customer | Posted 12/14/2021
    PBMCs either left resting or stimulated with LPS (1ug, 3h). Addition of GolgiPlug to detect intracellular IL1B production. Use of a secondary anti-goat antibody, conjugated to AF488.
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Tina Yuan
    Application: Immunohistochemistry
    Sample Tested: mouse oral tissue
    Species: Mouse
    Verified Customer | Posted 11/23/2020
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: THP-1 human acute monocytic leukemia cell line
    Species: Human
    Verified Customer | Posted 11/19/2020
    Human-derived THP-1 cells either left unstimulated or treated with LPS+ATP for 3 hours. Intracellular detection.
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Miguel Lopez Cuina
    Application: Western Blot
    Sample Tested: Brain (caudate putamen)
    Species: Mouse
    Verified Customer | Posted 05/23/2019
    I am somewhat disappointed. My lab published 4 years ago an article using this Ab. I boight it last september. There is no way I can get the bands ourselves and other authors have published: the proIL1-beta and the mature form, at around 35 and 17kDa. Attached is a representative blot of the many I have run. I have tried homemade 12% gels, BioRad 12 and 4-20% precast gels. I have tried both types of membranes (PVDF and Nitro 0.2), blocking with milk and BSA, different transfer times (35min 120V, 45min 100V, 1h 100V, 1.5h 100V), different Ab dilutions. And I still get nothing that comes to close to the pro-IL-1beta, and of course, the mature form is nowhere to be found. I do not wish to buy another brand, I believe this to be a good Ab, but since it is polyclonal, it might give different results depending on the lot. ***Bio-Techne Response: Thank you for reviewing our product. We are sorry to hear that this antibody did not perform as expected. We have been in touch with the customer to resolve this issue according to our Product Guarantee and to the customer’s satisfaction.***
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: IC-21 mouse macrophage cell line
    Species: Mouse
    Verified Customer | Posted 01/28/2019
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anthony Morales
    Application: Western Blot
    Sample Tested: IC-21 mouse macrophage cell line
    Species: Mouse
    Verified Customer | Posted 01/28/2019
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: CARLA RAMON
    Application: Immunohistochemistry
    Sample Tested: Brain tissue
    Species: Mouse
    Verified Customer | Posted 08/08/2018
    10.1093/cercor/bhy164
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Simple Western
    Sample Tested: J774A.1 mouse reticulum cell sarcoma macrophage cell line
    Species: Mouse
    Verified Customer | Posted 07/17/2018
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Paloma Narros
    Application: Western Blot
    Sample Tested: Cell Lysates and Macrophages Cell Supernatant
    Species: Mouse and Mouse macrophages
    Verified Customer | Posted 07/17/2018
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell Lysates
    Species: Mouse
    Verified Customer | Posted 07/17/2018
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Alisha Freeman
    Application: ELISA
    Sample Tested: A549 human lung carcinoma cell line
    Species: Human
    Verified Customer | Posted 05/17/2018
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Rupal Soder
    Application: Western Blot
    Sample Tested: IC-21 mouse macrophage cell line
    Species: Mouse
    Verified Customer | Posted 03/22/2018
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: bone marrow derived macrophage
    Species: Mouse
    Verified Customer | Posted 10/31/2017
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: L-929 mouse fibroblast cell line
    Species: Mouse
    Verified Customer | Posted 09/08/2017
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Michelle Chen
    Application: Block/Neutralize
    Sample Tested: Adult splenocytes
    Species: Mouse
    Verified Customer | Posted 04/26/2017
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Goutham Pattabiraman
    Application: Western Blot
    Sample Tested: Bone marrow cells, Serum-free Cell Culture Supernates and Cell culture supernatant
    Species: Mouse
    Verified Customer | Posted 04/25/2017
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Mouse IL-1 beta /IL-1F2 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: bone marrow derived macrophages
    Species: Mouse
    Verified Customer | Posted 02/14/2017
    Mouse IL‑1 beta/IL‑1F2 Antibody AF-401-NA
  • Name: Anonymous -
    Application: Western Blot
    Sample Tested: See PMID 23160153
    Species: Mouse
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous -
    Application: Immunocytochemistry
    Sample Tested: See PMID 23920123
    Species: Mouse
    Verified Customer | Posted 01/08/2015
  • Name: Anonymous -
    Application: ELISA
    Sample Tested: See PMID 21709053
    Species: Mouse
    Verified Customer | Posted 01/08/2015

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FAQs for Mouse IL‑1 beta/IL‑1F2 Antibody

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  • Q: Does catalog # AF-401-NA detect the pro or mature form of IL-1 beta in western blot?

    A: This would depend on the sample being used. The pro-form of the protein also contains the sequence for the mature form.  When cell lysates are used as the sample, the pro-form is mainly detected.  When cell culture supernatants containing mainly the mature form of the protein is used, only the mature form is detected.  This is exemplified in the following paper: J Immunol.(2015) 195, 1233.  http://www.jimmunol.org/content/195/3/1233/tab-figures-data

  • Q: Does Mouse IL-1 beta /IL-1F2 Antibody, Catalog # AF-401-NA, detect cleaved IL-1beta (17kDa)?

    A: The immunogen that was used to produce Catalog # AF-401-NA consists of the mature form of IL-1beta, amino acids Val118-Ser269. Since the pro-form also includes this sequence, we expect this antibody to detect the pro-form and cleaved IL-1beta.

Showing  1 - 2 of 2 FAQs Showing All

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