Phospho-specific Antibodies

Phosphorylation site-specific antibodies enable the analysis of key targets in normal and disease states, including cardiovascular disease, cancer, inflammation, neurodegenerative diseases, and diabetes. They are affinity-purified rabbit polyclonal and monoclonal antibodies that are monospecific for a target protein that is phosphorylated on a specific tyrosine, threonine, or serine residue. These tools are providing critical insight into the investigation of complex signal transduction events, including unraveling the intricacies of drug intervention in disease. Invitrogen phosphospecific antibodies are designed to dependably detect the key phosphospecific targets. Each antibody is validated for use in various applications.

Key phosphospecific antibody targets include

Applications

Invitrogen phosphospecific antibodies are designed to dependably detect phosphospecific targets in various applications.

Immunofluorescent analysis of Phospho-FAK (Try397)

Immunofluorescent analysis of of Phospho-FAK (Tyr397). Visualizing focal adhesion kinase with FAK (pY397) Polyclonal Antibody (Cat. No. 44-624G). HeLa cells were fixed and permeabilized using ice-cold 95% methanol. FAK (pY397) PSSA was labeled with Alexa Fluor 488 dye (green fluorescence, Zenon Alexa Fluor Rabbit IgG Labeling Kit, Cat. No. Z-25302), and nuclei were counterstained with DAPI (blue fluorescence, Cat. No. D21490). The image was captured using a Zeiss Axioplan 2 fluorescence microscope fitted with a 100× oil immersion lens.

Immunohistrochemistry analysis of ERK1/2 (pTpY185/187)

Immunohistochemistry analysis of ERK1/2 (pTpY185/187) showing staining in the cytoplasm and nucleus of paraffin-embedded human colon carcinoma tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with a ERK1/2 (pTpY185/187) polyclonal antibody (Cat. No. 44-680G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

ChIP- qPCR analysis of STAT1 (pTyr701) was performed with 5 µg of the STAT1 (pTyr701) Mouse monoclonal antibody (ST1P-11A5) (Cat. No. 33-3400) on sheared chromatin from 2 million HeLa cells treated with 50 ng/mL of IFN Gamma for one hour using the MAGnify Chromatin Immunoprecipitation System (Cat. No. 49-2024). Normal Mouse IgG was used as a negative IP control. The purified DNA from each ChIP sample was analyzed by StepOnePlus Real-Time PCR System (Cat. No. 4376600) with primers for the promoter of active GLS1 gene, used as positive control target, and the inactive SAT2, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

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