PA5-30612
antibody from Invitrogen Antibodies
Targeting: GMNN
Gem
Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- PA5-30612 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GMNN Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- 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.
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
Nature cell biology 2020 Jan;22(1):97-107
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using GMNN Polyclonal Antibody (Product # PA5-30612). Sample (30 µg of whole cell lysate). Lane A: HCT116. 12% SDS PAGE. GMNN Polyclonal Antibody (Product # PA5-30612) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-GMNN Polyclonal Antibody (Product # PA5-30612) and 30 kDa band corresponding to GMNN was enhanced upon double thymidine block followed by serum release and was also enhanced upon Nocodazole treatment. Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), HeLa treated with double thymidine block (for 18hr) (Lane 2), HeLa treated with double thymidine block (for 18hr) followed by serum release (for 8hr) (Lane 3), HeLa treated with Nocodazole (100nM for 2hr) (Lane 4), MOLT-4 (Lane 5) and K-562 (Lane 6) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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). .
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.
- 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
- 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.
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.