Invitrogen

Ghrelin Polyclonal Antibody

Advanced Verification

This Antibody was verified by Relative expression to ensure that the antibody binds to the antigen stated.

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Ghrelin Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of GHRL was performed using 70% confluent log phase MKN-45 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Ghrelin Rabbit Polyclonal Antibody (Product # PA1-1070) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Ghrelin Antibody in Western Blot (WB) — Western blot was performed using Anti-Ghrelin Polyclonal Antibody (Product # PA1-1070) and a 16 kDa band corresponding to Ghrelin was observed in LNCaP but not in DU 145. Whole cell extracts (30 µg lysate) of DU 145 (Lane 1), LNCaP (Lane 2) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Ghrelin Antibody — Antibody specificity was demonstrated by detection of differential basal expression of the target across DU 145, LNCaP owing to their inherent genetic constitution. Relative expression of Ghrelin was observed in LNCaP as compared to DU 145 using Anti-Ghrelin Polyclonal Antibody (Product # PA1-1070) in Western Blot. {RE}

Product Details

PA1-1070

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000	-
Immunocytochemistry (ICC/IF)	2 µg/mL	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Synthetic peptide corresponding to residues F(1) N A P F D V G I K L S G A Q Y Q Q H G R A L(23) of the rat Obestatin gene.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_1074594

Product Specific Information

PA1-1070 detects Obestatin in mouse and rat samples. This product does not detect endogenous levels of the hormone in rat stomach.

PA1-1070 has been successfully used in Western blot procedures. By Western blot, this antibody detects a ~2.5 kDa protein representing synthetic Obestatin.

The PA1-1070 immunogen is a synthetic peptide corresponding to residues F(1) N A P F D V G I K L S G A Q Y Q Q H G R A L(23) of the rat Obestatin gene. This sequence is conserved in mouse and only one amino acid residue difference in human. This peptide (Cat. # PEP-290) is available for use in neutralization and control experiments.

Target Information

GHRL encodes ghrelin-obestatin preproprotein, which generates ghrelin and obestatin. Ghrelin is an the ligand for the growth hormone secretagogue receptor type 1 and is involved in regulating growth hormone release. Ghrelin has an appeptite-stimulating effect and induces adiposity and stimulates gastric acid secretion. Obestatin may be the ligand for the orphan G protein-coupled receptor GPR39 and is involved in satiety and can have an appetite-reducing effect resulting in decreased food intake.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: AMCBX1; Appetite-regulating hormone; FIP3; Fip3p; Ghrelin; ghrelin 27; ghrelin 28; ghrelin, growth hormone secretagogue receptor ligand; Ghrelin-27; Ghrelin-28; ghrelin/obestatin; ghrelin/obestatin preprohormone; Growth hormone secretagogue; Growth hormone-releasing peptide; IKK-gamma; In2c-preproghrelin; IP1; IP2; IPD2; Motilin-related peptide; Obestatin; Obestatin-13; Obestatin-23; OTTHUMP00000207794; OTTHUMP00000207795; OTTHUMP00000207796; OTTHUMP00000207798; prepro-appetite regulatory hormone; preproghrelin; proghrelin; Protein M46

Gene Aliases: 2210006E23Rik; Ghr; GHRL; m46; MTLRP; MTLRPAP; UNQ524/PRO1066

UniProt ID: (Human) Q9UBU3, (Rat) Q9QYH7, (Mouse) Q9EQX0

Entrez Gene ID: (Human) 51738, (Rat) 59301, (Mouse) 58991

Molecular Function: peptide hormone