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PACAP (6-38), human, ovine, rat 

Cat. No.: HY-P0220
Handling Instructions

PACAP (6-38), human, ovine, rat is a potent PACAP receptor antagonist with IC50s of 30, 600, and 40 nM for PACAP type I receptor, PACAP type II receptor VIP1, and PACAP type II receptor VIP2, respectively.

For research use only. We do not sell to patients.

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PACAP (6-38), human, ovine, rat Chemical Structure

PACAP (6-38), human, ovine, rat Chemical Structure

CAS No. : 143748-18-9

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Description

PACAP (6-38), human, ovine, rat is a potent PACAP receptor antagonist with IC50s of 30, 600, and 40 nM for PACAP type I receptor, PACAP type II receptor VIP1, and PACAP type II receptor VIP2, respectively.

IC50 & Target

IC50: 30 nM (PACAP type I receptor), 600 nM (PACAP type II receptor VIP1), 40 nM (PACAP type II receptor VIP2)[1]

In Vitro

An increase of dopamine (DA) content by HPLC analysis and/or cell proliferation identified by MTT assay by Dexamethasone (DEX) is also observed which can be inhibited by PACAP (6-38) at concentration sufficient to block PACAP type 1 (PAC1) receptor. Pretreatment with PAC1 receptor antagonist PACAP (6-38) at 0.1 or 1 μM for 2 h significantly blocks this increase of DA content by 1 μM DEX. The MTT assay shows that DEX increases cell proliferation. Moreover, this action is also inhibited by the pre-incubation of PACAP (6-38). PACAP (6-38) at 1μM shows no effect on DA content and cell proliferation for 24 h. However, PACAP (6-38) at 0.3 μM has been mentioned to reduce the spontaneous tyrosine hydroxylase (TH) accumulation in differentiated retinal cultured cells for 5 days[2].

In Vivo

Intravesical administration of the PAC1 receptor antagonist, PACAP (6-38) (300 nM), significantly (p≤0.01) increases intercontraction interval (2.0-fold) and void volume (2.5-fold) in NGF-OE mice. Intravesical instillation of PACAP (6-38) also decreases baseline bladder pressure in NGF-OE mice. Intravesical administration of PACAP (6-38) (300 nM) significantly (p≤0.01) reduces pelvic sensitivity in NGF-OE mice but is without effect in WT mice[3].

Molecular Weight

4024.74

Formula

C₁₈₂H₃₀₀N₅₆O₄₅S

CAS No.

143748-18-9

Sequence

Phe-Thr-Asp-Ser-Tyr-Ser-Arg-Tyr-Arg-Lys-Gln-Met-Ala-Val-Lys-Lys-Tyr-Leu-Ala-Ala-Val-Leu-Gly-Lys-Arg-Tyr-Lys-Gln-Arg-Val-Lys-Asn-Lys-NH2

Sequence Shortening

FTDSYSRYRKQMAVKKYLAAVLGKRYKQRVKNK-NH2

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Solvent & Solubility
In Vitro: 

H2O

Peptide Solubility and Storage Guidelines:

1.  Calculate the length of the peptide.

2.  Calculate the overall charge of the entire peptide according to the following table:

  Contents Assign value
Acidic amino acid Asp (D), Glu (E), and the C-terminal -COOH. -1
Basic amino acid Arg (R), Lys (K), His (H), and the N-terminal -NH2 +1
Neutral amino acid Gly (G), Ala (A), Leu (L), Ile (I), Val (V), Cys (C), Met (M), Thr (T), Ser (S), Phe (F), Tyr (Y), Trp (W), Pro (P), Asn (N), Gln (Q) 0

3.  Recommended solution:

Overall charge of peptide Details
Negative (<0) 1.  Try to dissolve the peptide in water first.
2.  If water fails, add NH4OH (<50 μL).
3.  If the peptide still does not dissolve, add DMSO (50-100 μL) to solubilize the peptide.
Positive (>0) 1.  Try to dissolve the peptide in water first.
2.  If water fails, try dissolving the peptide in a 10%-30% acetic acid solution.
3.  If the peptide still does not dissolve, try dissolving the peptide in a small amount of DMSO.
Zero (=0) 1.  Try to dissolve the peptide in organic solvent (acetonitrile, methanol, etc.) first.
2.  For very hydrophobic peptides, try dissolving the peptide in a small amount of DMSO, and then dilute the solution with water to the desired concentration.
References
Cell Assay
[2]

PC12 cells (5×104 cells per well) are deposited in a 96-well flat-bottom culture plate. Cells are incubated with PACAP(6-38) (0.1 and 1.0 μM) for 2 h before the addition of Dexamethasone (DEX, 1 μM). Cells are harvested at 24 h later of treatment. At regular intervals after the additional treatments, 100 μL of 0.2 mg/mL MTT is added per each well, and cells are incubated for 3 h at 37°C. After incubation, the MTT reagent is discarded, and 100 μL of DMSO is then added. The experiment is performed at room temperature for 20 min. The absorbance is then measured with SPECTRA max 340PC ELISA reader at the wavelength of 570 nm[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Administration
[3]

Mice[3]
Two groups of mice are evaluated: WT mice receiving intravesical administration of vehicle (0.9% saline) and PACAP (6-38) (300 nM) (n=8) and NGF-OE mice receiving intravesical administration of vehicle (0.9% saline) and PACAP (6-38) (300 nM) (n=8). For intravesical administration of PACAP (6-38), mice are anesthetized with 2% isoflurane and PACAP (6-38) (<1.0 mL) is injected through the bladder catheter; the animals are maintained under anesthesia to prevent expulsion of PACAP (6-38) via a voiding reflex. In this procedure, PACAP (6-38) remains in the bladder for 30 min at which time, the drug is drained, the bladder washed with saline and animals recover from anesthesia for 20 min before experimentation[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
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Keywords:

PACAP (6-38), human, ovine, ratOthersInhibitorinhibitorinhibit

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PACAP (6-38), human, ovine, rat
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