- PA5-24822 - Invitrogen Antibodies H2AC11 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of HIST1H2AG 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 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HIST1H2AG Rabbit Polyclonal Antibody (Product # PA5-24822) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree 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 nuclear localization. Panel e represents control 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 HIST1H2AG 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 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HIST1H2AG Rabbit Polyclonal Antibody (Product # PA5-24822) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree 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 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 nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of HIST1H2AG in formalin fixed and paraffin embedded human lung carcinoma. Samples were incubated with HIST1H2AG polyclonal antibody (Product # PA5-24822) followed by peroxidase conjugation of the secondary antibody and DAB staining. This data demonstrates the use of this antibody for immunohistochemistry. Clinical relevance has not been evaluated.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry analysis of HepG2 cells using a HIST1H2AL polyclonal antibody (Product # PA5-24822) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry of HIST1H2AG in HepG2 cells (right histogram). Samples were incubated with HIST1H2AG polyclonal antibody (Product # PA5-24822) followed by FITC-conjugated goat-anti-rabbit secondary antibody. Negative control cell (left histogram).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Enrichment of endogenous HIST1H2AG protein at specific gene loci using Anti-HIST1H2AG Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-HIST1H2AG Rabbit Polyclonal Antibody (Product # PA5-24822, 3 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 for the promoters of c-Fos, ALDOA,GAPDH used as positive and MYOD, SAT2 satellite repeats, SAT Alpha used as negative target genes/binding sites. 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 HIST1H2AG protein at specific gene loci using Anti-HIST1H2AG Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-HIST1H2AG Rabbit Polyclonal Antibody (Product # PA5-24822, 3 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 for the promoters of c-Fos, ALDOA,GAPDH used as positive and MYOD, SAT2 satellite repeats, SAT Alpha used as negative target genes/binding sites. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

PA5-24822