IL-1 beta Recombinant Mouse Monoclonal Antibody [A7F7-R]
cat.: HA601036
Product Type: Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, ELISA, IHC-P
Clonality: Monoclonal
Clone number: A7F7-R
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 31 kDa
Isotype: IgG1
Immunogen: Recombinant protein within Human IL-1 beta aa 117-269 / 269.
Positive control: THP-1 treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours cell lysate, human tonsil tissue.
Subcellular location: Extracellular exosome, Secreted, Lysosome, Cytosol.
Recommended Dilutions:
  WB
  ELISA
  IHC-P

1:1,000-1:2,000
1:20,000
1:100
Uniprot #: SwissProt: P01584 Human | P10749 Mouse | Q63264 Rat
Alternative names: Catabolin H1 IFN beta inducing factor IL 1 IL 1 beta IL-1 beta IL1 IL1 BETA IL1B IL1B_HUMAN IL1F2 Interleukin 1 beta Interleukin 1 beta precursor interleukin 1, beta Interleukin-1 beta OAF Osteoclast activating factor OTTHUMP00000162031 Preinterleukin 1 beta Preinterleukin beta Pro interleukin 1 beta
Images
HA601036_1.jpg Fig1: Western blot analysis of IL-1 beta on different lysates with Mouse anti-IL-1 beta antibody (HA601036) at 1/1,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 31 kDa
Observed band size: 31 kDa

Exposure time: 1 minute;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601036) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HA601036_2.jpg Fig2: Western blot analysis of IL-1 beta on different proteins with Mouse anti-IL-1 beta antibody (HA601036) at 1/2,000 dilution.

Lane 1: Recombinant mouse IL-1 beta
Lane 2: Recombinant rat IL-1 beta
Lane 3: Recombinant human IL-1 beta

Lysates/proteins at 50 ng/Lane.

Exposure time: 30 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601036) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HA601036_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-IL-1 beta antibody (HA601036) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601036) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA601036_4.jpg Fig4: IL-1 beta Antibody (HA601036) in indirect ELISA.

Indirect ELISA analysis of IL-1 beta was performed by coating wells of a 96-well plate with 50 µl per well of IL-1 beta antigen diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 50 µl per well of a mouse IL-1 beta monoclonal antibody starting at a concentration of 20 µg/mL and serially diluting it to a concentration of 1.28 ng/mL for 2 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-mouse IgG secondary antibody at a dilution of 1:10,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 5 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.