IL-10 Recombinant Rabbit Monoclonal Antibody [JE59-71]
cat.: HA722032
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
Images
HA722032_1.jpg 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.
HA722032_2.jpg 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.
HA722032_3.jpg 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.