Anti-CD83 antibody [EPR23809-19]

• Flow Cyt

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Flow Cytometry - Anti-CD83 antibody [EPR23809-19] (AB275021)

Flow cytometric analysis of HDLM-2 (Human Hodgkin lymphoma) cells labelling CD83 with ab275021 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD83 antibody [EPR23809-19] (AB275021)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD83 with ab275021 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil. The section was incubated with ab275021 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt

Lab

Flow Cytometry - Anti-CD83 antibody [EPR23809-19] (AB275021)

Flow cytometry staining of human monocyte-derived dendritic cells (top) or human monocyte-derived dendritic cells treated with LPS, 24 Hours, 100ng/mL (bottom), with ab275021 (right) or Rabbit IgG (monoclonal) Phycoerythrin (ab209478) (left). Cells were incubated for 30 mins on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab275021 or Rabbit IgG (monoclonal) Phycoerythrin (ab209478) (1x 10⁶ in 100 µl at 0.2 μg/ml (1/2,500 dilution)) for 30 mins on ice. The cells were simultaneously stained with CD209.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilutionfor 30 mins on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• IP

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Immunoprecipitation - Anti-CD83 antibody [EPR23809-19] (AB275021)

CD83 was immunoprecipitated from 0.35 mg HDLM-2 (human hodgkin lymphoma) whole cell lysate with ab275021 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275021 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HDLM-2 whole cell lysate 10 μg.

Lane 2 : ab275021 IP in HDLM-2 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275021 in HDLM-2 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 145 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 15320871).

All lanes:

Immunoprecipitation - Anti-CD83 antibody [EPR23809-19] (ab275021)

Predicted band size: 23 kDa

Observed band size: 40-50 kDa

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• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD83 antibody [EPR23809-19] (AB275021)

Immunohistochemical analysis of paraffin-embedded human Hodgkin's lymphoma tissue labeling CD83 with ab275021 at 1/500 dilution followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human Hodgkin's lymphoma (PMID : 24633193). The section was incubated with ab275021 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.