MA5-16358

antibody from Invitrogen Antibodies

Targeting: ANO1 DOG1, FLJ10261, ORAOV2, TAOS2, TMEM16A

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-16358

Provider

Invitrogen Antibodies

Product name

DOG-1 Recombinant Rabbit Monoclonal Antibody (SP31)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is gastrointestinal stromal tumor.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

SP31

Vial size

500 µL

Concentration

0.055 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

TMEM16A/ANO1 Inhibits Apoptosis Via Downregulation of Bim Expression.

Godse NR, Khan N, Yochum ZA, Gomez-Casal R, Kemp C, Shiwarski DJ, Seethala RS, Kulich S, Seshadri M, Burns TF, Duvvuri U
Clinical cancer research : an official journal of the American Association for Cancer Research 2017 Dec 1;23(23):7324-7332

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TMEM16A/ANO1 is differentially expressed in HPV-negative versus HPV-positive head and neck squamous cell carcinoma through promoter methylation.

Dixit R, Kemp C, Kulich S, Seethala R, Chiosea S, Ling S, Ha PK, Duvvuri U
Scientific reports 2015 Nov 13;5:16657

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Plexiform fibromyxoma (plexiform angiomyxoid myofibroblastic tumor) of stomach: an unusual presentation as a fistulating abscess.

Lee PW, Yau DT, Lau PP, Chan JK
International journal of surgical pathology 2014 May;22(3):286-90

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Na+/Ca2+ exchangers regulate the migration and proliferation of human gastric myofibroblasts.

Kemény LV, Schnúr A, Czepán M, Rakonczay Z Jr, Gál E, Lonovics J, Lázár G, Simonka Z, Venglovecz V, Maléth J, Judák L, Németh IB, Szabó K, Almássy J, Virág L, Geisz A, Tiszlavicz L, Yule DI, Wittmann T, Varró A, Hegyi P
American journal of physiology. Gastrointestinal and liver physiology 2013 Oct 15;305(8):G552-63

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Contribution of DOG1 expression to the diagnosis of gastrointestinal stromal tumors.

Kara T, Serinsoz E, Arpaci RB, Gubur O, Orekici G, Ata A, Colak T, Arican A
Pathology, research and practice 2013 Jul;209(7):413-7

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-DOG1 Monoclonal antibody(SP31) (Product # MA5-16358) and a 110 kDa band corresponding to DOG1 was observed in PC-3, HT-29, HCT 116 and COLO 205 but not in DU 145 and HEK 293 which are reported to be negative. Membrane enriched extracts (30 µg lysate) of PC-3 (Lane 1), HT-29 (Lane 2), DU 145 (Lane 3), HCT 116 (Lane 4), COLO 205 (Lane 5) and HEK 293 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of DOG-1 was performed using 70% confluent log phase PC-3 and DU 145 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. PC-3 cells were labeled with DOG-1 Monoclonal Antibody (Product # MA5-16358) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight 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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image of PC-3 showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. Panel f represents the merged image of DU 145 cells showing no expression for DOG-1 protein. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of DOG-1 using anti-DOG-1 Monoclonal Antibody (Product # MA5-16358) in GIST Cancer Tissue. The recommened dilution for this antibody in immunohistochemistry applications is 1:100.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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