WB result of P4HB Recombinant Rabbit mAb
Primary antibody: P4HB Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: HeLa whole cell lysate 20 µg
Lane 2: A431 whole cell lysate 20 µg
Lane 3: HepG2 whole cell lysate 20 µg
Lane 4: HL-60 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 57 kDa
Observed MW: 60 kDa
P4HB Recombinant Rabbit mAb (S-1524-120)
P4HB Recombinant Rabbit mAb (S-1524-120)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | P4HB |
| Synonyms | Protein disulfide-isomerase; PDI; Cellular thyroid hormone-binding protein; Prolyl 4-hydroxylase subunit beta; p55; ERBA2L; PDI; PDIA1; PO4DB |
| Immunogen | Synthetic Peptide |
| Location | Cell membrane, Endoplasmic reticulum |
| Accession | P07237 |
| Clone Number | S-1524-120 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ICFCM |
| Reactivity | Hu |
| Positive Sample | HeLa, A431, HepG2, HL-60 |
| Predicted Reactivity | JaMq, Or |
| 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 | 1:1000 | Hu |
| ICC | 1:500 | Hu |
| ICFCM | 1:500 | Hu |
Background
P4HB protein, also known as prolyl 4-hydroxylase subunit beta, is a multifunctional protein that belongs to the protein disulfide isomerase family. It serves as an oxidoreductase, isomerase, and molecular chaperone, and is involved in the hydroxylation of prolyl residues in preprocollagen, the formation, breakage, and rearrangement of disulfide bonds, and the inhibition of aggregation of misfolded proteins. P4HB is found in various cellular compartments, including the endoplasmic reticulum, mitochondria, cytosol, and the cell surface. It also plays a role in the regulation of reactive oxygen species (ROS) generation, prevention of ER stress, and protein misfolding. Additionally, P4HB has been implicated in several types of cancer and may be associated with immune responses, as it strongly co-expresses with immune-related genes and is involved in antigen processing and presentation, as well as protein processing in the endoplasmic reticulum. High expression levels of P4HB are correlated with worse prognosis in various cancers, suggesting its potential as a diagnostic biomarker and therapeutic target.
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Western Blot
FC
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) labelling P4HB antibody at 1/500 dilution (0.1 μg)/ (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.
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human cerebral cortex. Anti-P4HB antibody was used at 1/1000 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-P4HB antibody was used at 1/1000 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-P4HB antibody was used at 1/1000 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-P4HB antibody was used at 1/1000 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 renal clear cell carcinoma. Anti-P4HB antibody was used at 1/1000 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 A431 cells. Anti- P4HB 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).
