| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | JE59-71 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 21 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human IL-10 aa 1-100. |
| Positive control: | THP-1 cell lysate, THP-1 treated with 800nM TPA overnight then treated with 100ng/mL LPS for 6 hours add 300ng/mL BFA for 3 hours cell lysate, mouse spleen tissue lysate, rat spleen tissue 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, rat spleen tissue. |
| Subcellular location: | Secreted. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:1,000 1:500 1:1,000 |
| Uniprot #: | SwissProt: P22301 Human | P18893 Mouse | P29456 Rat |
| Alternative names: | CSIF Cytokine synthesis inhibitory factor GVHDS IL 10 IL-10 IL10 IL10_HUMAN IL10A Interleukin 10 Interleukin-10 MGC126450 MGC126451 T-cell growth inhibitory factor TGIF |
|
Fig1:
Western blot analysis of IL-10 on different lysates with Rabbit anti-IL-10 antibody (HA722032) at 1/1,000 dilution. Lane 1: THP-1 cell lysate (30 µg/Lane) 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 (30 µg/Lane) Lane 3: Mouse spleen tissue lysate (40 µg/Lane) Lane 4: Rat spleen tissue lysate (40 µg/Lane) Predicted band size: 21 kDa Observed band size: 21 kDa Exposure time: 1 minutes 59 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 (HA722032) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
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-10 with Rabbit anti-IL-10 antibody (HA722032) at 1/500 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 Rabbit anti-IL-10 antibody (HA722032) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
|
Fig3:
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-IL-10 antibody (HA722032) at 1/1,000 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 (HA722032) at 1/1,000 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. |