PA5-30612

antibody from Invitrogen Antibodies

Targeting: GMNN Gem

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-30612

Provider

Invitrogen Antibodies

Product name

GMNN Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Recommended positive controls: HCT116. Predicted reactivity: Rhesus Monkey (94%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

p120-catenin-dependent collective brain infiltration by glioma cell networks.

Gritsenko PG, Atlasy N, Dieteren CEJ, Navis AC, Venhuizen JH, Veelken C, Schubert D, Acker-Palmer A, Westerman BA, Wurdinger T, Leenders W, Wesseling P, Stunnenberg HG, Friedl P
Nature cell biology 2020 Jan;22(1):97-107

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Geminin in paraformaldehyde-fixed HeLa cells using a Geminin polyclonal antibody (Product # PA5-30612) at a 1:500 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of GMNN was performed using 70% confluent log phase HeLa 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. The cells were labeled with GMNN Polyclonal Antibody (Product # PA5-30612) 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, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents cells with no primary antibody to assess background. The images were captured at 60X magnification..

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of GMNN was performed using 70% confluent log phase HeLa 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. The cells were labeled with GMNN Polyclonal Antibody (Product # PA5-30612) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (Heavy Chain) 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, 1:300). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents cells with no primary antibody to assess background. The images were captured at 60X magnification..

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

GMNN Polyclonal Antibody detects Geminin protein at nucleus on human breast cancer by immunohistochemical analysis. Sample: Paraffin-embedded breast cancer. GMNN Polyclonal Antibody (Product # PA5-30612) dilution: 1:250. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Enrichment of endogenous GMNN protein at specific gene loci using Anti-GMNN Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-GMNN Rabbit Polyclonal Antibody (Product # PA5-30612, 5 µg) on sheared chromatin from 2 million Hela cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over cFOS, GAPDH and SAT2. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Enrichment of endogenous GMNN protein at specific gene loci using Anti-GMNN Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-GMNN Rabbit Polyclonal Antibody (Product # PA5-30612, 5 µg) on sheared chromatin from 2 million Hela cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over cFOS, GAPDH and SAT2. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.