WB result of GRP75 Rabbit pAb
Primary antibody: GRP75 Rabbit pAb at 1/1000 dilution
Lane 1: MCF7 whole cell lysate 20 µg
Lane 2: HeLa whole cell lysate 20 µg
Lane 3: PANC-1 whole cell lysate 20 µg
Lane 4: Jurkat whole cell lysate 20 µg
Lane 5: K562 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 74 kDa
Observed MW: 74 kDa
GRP75 Rabbit Polyclonal Antibody
GRP75 Rabbit Polyclonal Antibody
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | GRP75 |
| Synonyms | Stress-70 protein, mitochondrial; 75 kDa glucose-regulated protein (GRP-75); Heat shock 70 kDa protein 9; Heat shock protein family A member 9; Mortalin (MOT); Peptide-binding protein 74 (PBP74); HSPA9B; mt-HSP70; HSPA9 |
| Immunogen | Recombinant Protein |
| Location | Cytoplasm, Nucleus, Mitochondrion, Endoplasmic reticulum |
| Accession | P38646 |
| Antibody Type | Polyclonal antibody |
| Isotype | IgG |
| Application | WB, IHC-P, ICC |
| Reactivity | Hu, Ms, Rt, Mk |
| Positive Sample | MCF7, HeLa, PANC-1, Jurkat, K562, NIH/3T3, mouse liver, PC-12, rat brain, COS-7 |
| Purification | Immunogen Affinity |
| 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, Ms, Rt, Mk |
| IHC-P | 1:200 | Hu, Ms, Rt |
| ICC | 1:500 | Hu |
Background
GRP75, also known as mortalin or HSPA9, is an evolutionarily highly-conserved 75-kDa heat-shock protein that resides mainly in mitochondria but also shuttles to cytosol, ER, plasma membrane and even nucleus; it functions as a multifunctional molecular chaperone that binds unfolded or misfolded proteins in an ATP-dependent manner to prevent aggregation, drives import and proper folding of nascent mitochondrial precursors, participates in mitochondrial biogenesis, iron–sulfur cluster assembly, calcium homeostasis, and quality-control proteolysis, and additionally regulates stress response, cell-cycle progression, senescence and apoptosis by modulating p53, telomerase and other signaling pathways; consequently, its overexpression is frequently observed in many tumors where it promotes survival and chemoresistance, whereas its loss-of-function mutations cause severe neurodegenerative and mitochondrial disorders, making GRP75 a pivotal hub linking bioenergetics, proteostasis and human disease.
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Western Blot
WB result of GRP75 Rabbit pAb
Primary antibody: GRP75 Rabbit pAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Lane 2: mouse liver lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 74 kDa
Observed MW: 74 kDa
WB result of GRP75 Rabbit pAb
Primary antibody: GRP75 Rabbit pAb at 1/1000 dilution
Lane 1: PC-12 whole cell lysate 20 µg
Lane 2: rat brain lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 74 kDa
Observed MW: 74 kDa
WB result of GRP75 Rabbit pAb
Primary antibody: GRP75 Rabbit pAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti- rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 74 kDa
Observed MW: 74 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human kidney. Anti-GRP75 antibody was used at 1/200 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 adenocarcinoma. Anti-GRP75 antibody was used at 1/200 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-GRP75 antibody was used at 1/200 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 mouse kidney. Anti-GRP75 antibody was used at 1/200 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 rat kidney. Anti-GRP75 antibody was used at 1/200 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 MCF7 cells. Anti-GRP75 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).
