WB result of BAG3 Recombinant Rabbit mAb
Primary antibody: BAG3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HL-60 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: K562 whole cell lysate 20 µg
Lane 4: 293T whole cell lysate 20 µg
Negative control: HL-60 whole cell lysate
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 62 kDa
Observed MW: 80 kDa
BAG3 Recombinant Rabbit mAb (S-1837-22)
BAG3 Recombinant Rabbit mAb (S-1837-22)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | BAG3 |
| Synonyms | BAG family molecular chaperone regulator 3; BAG-3; Bcl-2-associated athanogene 3; Bcl-2-binding protein Bis; Docking protein CAIR-1; BIS |
| Immunogen | Recombinant Protein |
| Location | Cytoplasm, Nucleus |
| Accession | O95817 |
| Clone Number | S-1837-22 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu |
| Positive Sample | HeLa, K562, 293T |
| 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 | 1:1000 | Hu |
| IHC-P | 1:250 | Hu |
| ICC | 1:500 | Hu |
Background
BAG3 protein, also known as Bcl2-associated athanogene 3, is a 575 amino acid multi-functional protein that is widely expressed across various tissues but is most abundant in cardiac muscle, skeletal muscle, the brain, and many cancers. It plays a crucial role in maintaining cellular homeostasis by supporting the autophagy of misfolded proteins and damaged organelles, inhibiting apoptosis, preserving mitochondrial homeostasis, and facilitating excitation-contraction coupling through the L-type calcium channel and the beta-adrenergic receptor. High levels of BAG3 are associated with resistance to chemotherapy in malignant cells, while both loss of function and gain of function variants are linked to cardiomyopathy. BAG3 is also characterized by its multiple protein-protein binding domains, which enable it to establish discrete intracellular micro-environments based on its specific responsibilities and locations within the cell. .
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Western Blot
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-BAG3 antibody was used at 1/250 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 cardiac muscle. Anti-BAG3 antibody was used at 1/250 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 hepatocellular carcinoma. Anti-BAG3 antibody was used at 1/250 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-BAG3 antibody was used at 1/250 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-BAG3 antibody was used at 1/250 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 HeLa cells. Anti- BAG3 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).
