Rabbit anti-CD276 Recombinant Monoclonal Antibody(249-26)

Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized LNCaP (Human prostate carcinoma epithelial cell) cells labelling B7-H3/CD276 antibody at 1/500 dilution (0.1 ug) / (red) compared with a Rabbit monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody.

ICC shows positive staining in LnCaP cells. Anti-B7-H3 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (red).

IHC shows positive staining in paraffin-embedded human placenta. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human colon. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

Negative control: IHC shows negative staining in paraffin-embedded human skeletal muscle. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human breast cancer. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human colon cancer. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human endometrial cancer. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human lung cancer. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

IHC shows positive staining in paraffin-embedded human ovarian cancer. Anti- B7-H3/CD276 antibody was used at 1/2000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.

B7-H3/CD276 Rabbit mAb at 1/50 dilution (1 ug) immunoprecipitating B7-H3/CD276 in 0.4 mg LnCap whole cell lysate. Western blot was performed on the immunoprecipitate using B7-H3/CD276 Rabbit mAb at 1/1000 dilution. Secondary antibody (HRP) for IP was used at 1/400 dilution. Lane 1: LnCap whole cell lysate 20 ug (Input) Lane 2: B7-H3/CD276 Rabbit mAb IP in LnCap whole cell lysate Lane 3: Rabbit monoclonal IgG IP in LnCap whole cell lysate Predicted MW: 90 kDa Observed MW: 90~110 kDa Exposure time: 60 s

WB result of B7-H3/CD276 Rabbit mAb Primary antibody: B7-H3/CD276 Rabbit mAb at 1/1000 dilution Lane 1: Raji whole cell lysate 20 ug Lane 2: LNCaP whole cell unboiled RIPA lysate 20 ug Lane 3: HEK-293 whole cell unboiled RIPA lysate 20 ug Lane 4: U-2 OS whole cell lysate 20 ug Negative control: Raji whole cell lysate Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution Predicted MW: 90 kDa Observed MW: 90~110 kDa Exposure time: 180 s