WB result of B7-H4 Rabbit mAb
Primary antibody: B7-H4 Rabbit mAb at 1/200 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: SK-BR-3 whole cell lysate 20 µg
Lane 3: T-47D whole cell lysate 20 µg
Negative control: HeLa whole cell lysate
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 31 kDa
Observed MW: 65, 45 kDa
(This blot was developed with high sensitivity substrate)
B7-H4 Recombinant Rabbit mAb (SDT-314-447)
B7-H4 Recombinant Rabbit mAb (SDT-314-447)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | B7-H4 |
| Synonyms | V-set domain-containing T-cell activation inhibitor 1, B7 homolog 4, B7h.5, Protein B7S1, T-cell costimulatory molecule B7x, VTCN1 |
| Immunogen | Recombinant Protein |
| Location | Cell membrane |
| Accession | Q7Z7D3 |
| Clone Number | SDT-314-447 |
| Antibody Type | Recombinant mAb |
| Application | WB, IHC-P, ICC, IP, IF |
| Reactivity | Hu |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05%BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:200 | |
| IP | 1:50 | |
| IHC | 1:400 | |
| ICC | 1:50 | |
| IF | 1:500 |
Background
B7-H4 is a transmembrane protein that binds an unknown receptor on activated T cells resulting in inhibition of T-cell effector function via cell cycle arrest, decreased proliferation, and reduced IL-2 production. B7-H4 is up-regulated on the surface of cancer cells and immunosuppressive tumor-associated macrophages (TAMs) in a variety of human cancers. Notably, B7-H4 expression levels inversely correlate with patient survival in ovarian cancer, cholangiocarcinoma, breast cancer and endometrial cancer, making B7-H4 an attractive candidate for therapeutic intervention.
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Western Blot
WB result of B7-H4 Rabbit mAb
Primary antibody: B7-H4 Rabbit mAb at 1/200 dilution
Lane 1: recombinant human B7-H1 protein 10ng (CAT.NO.S0A6006)
Lane 2: recombinant human B7-H2 protein 10ng (CAT.NO.UA010058)
Lane 3: recombinant human B7-H3 protein 10ng (CAT.NO.UA010010)
Lane 4: recombinant human B7-H4 protein 10ng (CAT.NO.UA010121)
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 31 kDa
Observed MW: 43~55 kDa
(This blot was developed with high sensitivity substrate)
IP
B7-H4 Rabbit mAb at 1/50 dilution (1 µg) immunoprecipitating B7-H4 in 0.4 mg SK-BR-3 whole cell lysate. Western blot was performed on the immunoprecipitate using B7-H4 Rabbit mAb at 1/1000 dilution. Secondary antibody (HRP) for IP was used at 1/400 dilution. Lane 1: SK-BR-3 whole cell lysate 20 µg (Input) Lane 2: B7-H4 Rabbit mAb IP in SK-BR-3 whole cell lysate Lane 3: Rabbit monoclonal IgG IP in SK-BR-3 whole cell lysate Predicted MW: 45 kDa Observed MW: 65 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human breast. Anti-B7-H4 antibody was used at 1/400 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-H4 antibody was used at 1/400 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-H4 antibody was used at 1/400 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-H4 antibody was used at 1/400 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-H4 antibody was used at 1/400 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.
Immunocytochemistry
ICC shows positive staining in SK-BR-3 cells. Anti-B7-H4 antibody was used at 1/50 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.
Negative control: ICC shows negative staining in HeLa cells. Anti-B7-H4 antibody was used at 1/50 dilution 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.
Immunofluorescence
IF shows positive staining in paraffin-embedded human breast cancer (case 1). Anti-B7-H4 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)(S0B4004) was used as secondary antibody at 1/500 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
IF shows positive staining in paraffin-embedded human breast cancer (case 2). Anti-B7-H4 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)(S0B4004) was used as secondary antibody at 1/500 dilution. Counterstained with DAPI (Blue). Heat mediated antigen retrieval with EDTA buffer pH9.0 was performed before commencing with IF staining protocol.
