Anti-CD83 antibody [EPR22405]

• Flow Cyt

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Flow Cytometry - Anti-CD83 antibody [EPR22405] (AB244204)

Flow cytometric analysis of HDLM-2 (human Hodgkin lymphoma) cell line labeling CD83 with ab244204 at 1/50 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD83 antibody [EPR22405] (AB244204)

Immunofluorescent analysis of HDLM-2 (human Hodgkin lymphoma) cells fixed with 100% methanol, labeling CD83 with ab244204 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in HDLM-2 cells is observed. We recommend using methanol instead of PFA fixation. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.

• Flow Cyt

Unknown

Flow Cytometry - Anti-CD83 antibody [EPR22405] (AB244204)

Flow cytometric analysis of human peripheral blood mononuclear cell (PBMC) labeling CD83 with ab244204 at 1/50 (Right panel) compared with a Rabbit monoclonal IgG (ab172730) (Left panel). Goat anti rabbit IgG (Dylight^® 488, ab98462), at 1/2000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left panel) or anti-CD83 RabMab (Right panel). Then stained with Alexa Fluor^® 647-conjugated anti-CD14. The result is consistent with what has been described in the literature. (PMID : 15320871).

Gated on viable cells.

• WB

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Western blot - Anti-CD83 antibody [EPR22405] (AB244204)

The molecular weight observed is consistent with what has been described in the literature (PMID : 15320871). The bands above 23 kDa are glycosylated forms.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-CD83 antibody [EPR22405] (ab244204) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 10 µg

Lane 2:

THP-1 treated with 100ng/ml LPS for 3 hours, whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 23 kDa

false

• WB

Unknown

Western blot - Anti-CD83 antibody [EPR22405] (AB244204)

The molecular weight observed is consistent with what has been described in the literature (PMID : 15320871). The bands above 23 kDa are glycosylated forms.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : 26 seconds.

All lanes:

Western blot - Anti-CD83 antibody [EPR22405] (ab244204) at 1/1000 dilution

Lane 1:

Untreated Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 2:

Raji whole cell lysate treated with PNGase F at 20 µg

Lane 3:

Untreated HDLM-2 (human hodgkin lymphoma) whole cell lysate at 20 µg

Lane 4:

HDLM-2 whole cell lysate treated with PNGase F at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 23 kDa

Observed band size: 40-50 kDa

false