SCTR Antibody, HRP conjugated
Description
Introduction to SCTR Antibody, HRP Conjugated
The SCTR Antibody, HRP conjugated is a specialized reagent combining a primary antibody targeting the Secretin Receptor (SCTR) protein with Horseradish Peroxidase (HRP), an enzyme enabling direct detection in assays like ELISA, immunohistochemistry (IHC), and Western Blot (WB). This conjugate eliminates the need for secondary antibodies, streamlining workflows in research and diagnostics .
Conjugation Process and Mechanism
The conjugation of HRP to SCTR antibodies involves covalent bonding using proprietary reagents, typically under near-neutral pH conditions to preserve antibody integrity. Key steps include:
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Buffer Preparation: Antibodies are diluted in amine-free buffers (e.g., HEPES, phosphate) to avoid interference with conjugation chemistry .
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Molar Ratios: Optimal HRP-to-antibody ratios (1:4 to 1:1) are critical for efficiency, balancing enzyme activity and antibody recovery .
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Stabilization: Post-conjugation, additives like LifeXtend™ protect HRP activity from degradation .
ELISA
Direct detection in sandwich ELISA eliminates secondary antibodies. For example:
Immunohistochemistry (IHC)
HRP conjugates enable chromogenic detection via DAB or ECL, with no cross-reactivity from secondary antibodies . Recommended dilutions range from 1:20 to 1:500 depending on tissue type .
Western Blot (WB)
Direct detection reduces background noise. Key dilutions:
| Supplier | Dilution Range | Applications |
|---|---|---|
| Cusabio | 1:500–1:5000 | WB, IHC |
| Proteintech | 1:5000–1:50000 | WB, IF/ICC |
| Abbexa | Optimal per assay | ELISA |
Product Details
Cross-Reactivity and Specificity
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Specificity: No significant cross-reactivity with analogues reported .
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Limitations: Sodium azide and primary amines inhibit HRP activity .
ELISA Performance
Immunofluorescence (IF) and IHC
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Proteintech’s SCTR Antibody shows strong staining in THP-1 cells and mouse pancreas .
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Abbexa’s HRP-conjugated antibody is validated for ELISA but lacks IHC data .
Comparative Analysis of Conjugation Kits
Product Specs
Target Background
The Secretin Receptor (SCTR) is a G protein-coupled receptor that binds secretin, a hormone involved in regulating duodenal pH, food intake, and water homeostasis. SCTR activation via G protein signaling stimulates adenylyl cyclase. Specifically, secretin binding modulates duodenal pH by inhibiting gastric acid secretion from parietal cells and stimulating bicarbonate production from pancreatic ductal cells. Beyond pH regulation, SCTR plays a crucial role in diet-induced thermogenesis, acting as a non-sympathetic brown adipose tissue (BAT) activator to mediate prandial thermogenesis and promote satiation. This occurs through secretin binding to SCTR in brown adipocytes, triggering lipolysis, a process sensed by the brain leading to satiety. SCTR also stimulates lipolysis in white adipocytes. Furthermore, it contributes significantly to cellular osmoregulation by influencing renal water reabsorption and plays a role in synaptic plasticity within the central nervous system.
The following research highlights the diverse roles and regulatory mechanisms of the SCTR gene:
- Elevated SCTR expression in primary sclerosing cholangitis liver samples compared to healthy controls. (PMID: 27115285)
- Structure and function of cross-class complexes of G protein-coupled Secretin and Angiotensin 1a Receptors. (PMID: 27330080)
- Identification of charge-charge interactions between secretin and SCTR residues using cysteine trapping. (PMID: 26740626)
- SCTR suppresses normal breast cell proliferation, while stimulating cancer cell proliferation and migration, with downregulation mediated by promoter methylation. (PMID: 26397240)
- High SCTR expression is associated with liver metastases of pancreatic neuroendocrine tumors. (PMID: 25241033)
- Secretin's potent modulation of adipocyte function, enhancing substrate cycling. (PMID: 22565418)
- Down-regulatory effects of NRSF on hSCTR gene expression through suppression of Sp1-mediated transactivation. (PMID: 23168245)
- Secretin and/or SCTR modulation as potential therapeutic tools in cholangiocarcinoma treatment. (PMID: 19904746)
- Increased SCTR expression competes for RAMP3 association with CLR, affecting adrenomedullin receptor function. (PMID: 19886671)
- Promoter methylation's role in regulating SCTR gene expression. (PMID: 14645499)
- Presence of SCTR transcripts in Purkinje and basket cells of the human cerebellum. (PMID: 15706223)
- Marked reduction in SCTR binding in ductal neoplasia. (PMID: 16192632)
- Secretin receptor oligomerization through transmembrane segment interactions. (PMID: 16819820)
- High SCTR expression in cholangiocarcinomas for in vivo targeting and differential diagnosis. (PMID: 16935383)
- Identification of a novel SCTR spliceoform in pancreatic and bile duct cancers, and its use in a diagnostic assay. (PMID: 17678920)
- SCTR and SCTR-variant expression in all gastrinomas. (PMID: 17711922)
- Secretin receptors as new markers for bronchopulmonary carcinoid tumors. (PMID: 18223557)
- Family B G-protein-coupled receptor oligomerization, including SCTR. (PMID: 18401761)
- Evidence suggesting SCTR exists as a structurally-specific homodimer, not higher-order oligomers. (PMID: 18680717)
