IL-1 beta Mouse Monoclonal Antibody [A7A10]
cat.: HA601035
| Product Type: | Mouse monoclonal IgG1, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, ELISA, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | A7A10 |
| 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, THP-1 cells treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours. |
| Subcellular location: | Extracellular exosome, Secreted, Lysosome, Cytosol. |
| Recommended Dilutions:
WB ELISA IF-Cell |
1:1,000-1:2,000 1:10,000 1:100 |
| Uniprot #: | SwissProt: P01584 Human |
| 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
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Fig1:
Western blot analysis of IL-1 beta on different lysates with Mouse anti-IL-1 beta antibody (HA601035) 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: 3 minutes; 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 (HA601035) 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. |
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Fig2:
Immunocytochemistry analysis of THP-1 cells treated with 80nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours labeling IL-1 beta with Mouse anti-IL-1 beta antibody (HA601035) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-IL-1 beta antibody (HA601035) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
IL-1 beta Antibody (HA601035) 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”.