PA3-116

antibody from Invitrogen Antibodies

Targeting: CCKAR

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-116

Provider

Invitrogen Antibodies

Product name

CCKAR Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA3-116 detects CCK1-Receptor in human samples. PA3-116 has been successfully used in Western blot, immunocytochemistry and immunohistochemistry procedures. By Western blot, this antibody detects an ~80-90 kDa protein representing CCK1-Receptor in human samples. In immunohistochemistry, PAC1 immunoreactivity is predominantly localized at the plasma membrane and in cells of the gastric mucosa, myenteric ganglion cells, myenteric nerve fibers in the gasrtointestinal tract. PA3-116 immunogen is a synthetic peptide, KLH conjugated, corresponding to the residues T(409) G A S L S R F S Y S H M S A S V P P Q (428) of human VPAC2 receptor.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

Cholecystokinin receptor antagonist halts progression of pancreatic cancer precursor lesions and fibrosis in mice.

Smith JP, Cooper TK, McGovern CO, Gilius EL, Zhong Q, Liao J, Molinolo AA, Gutkind JS, Matters GL
Pancreas 2014 Oct;43(7):1050-9

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Cholecystokinin mediates progression and metastasis of pancreatic cancer associated with dietary fat.

Matters GL, Cooper TK, McGovern CO, Gilius EL, Liao J, Barth BM, Kester M, Smith JP
Digestive diseases and sciences 2014 Jun;59(6):1180-91

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Immunohistochemical localization of CCK1 cholecystokinin receptors in normal and neoplastic human tissues.

Schulz S, Röcken C, Mawrin C, Schulz S
The Journal of clinical endocrinology and metabolism 2005 Nov;90(11):6149-55

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Pancreas (Lane 1) and Rat Pancreas (Lane 2). The blot was probed with Anti-CCKAR Rabbit Polyclonal Antibody (Product # PA3-116, 1:1,000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 47 kDa band corresponding to CCKAR was observed across the tissues tested. An additional band around 80 kDa was observed in Rat Pancreas which corresponds to another isoform of CCKAR. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CCK-A Receptor was performed using 70% confluent log phase PANC-1 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 CCKAR Rabbit Polyclonal Antibody (Product # PA3-116) at 1:250 dilution 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) at 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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of CCK1-Receptor in human gastric mucosa samples using Product # PA3-116.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL