MA5-14999

antibody from Invitrogen Antibodies

Targeting: AKT2 PKBβ

Western blot (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14999

Provider

Invitrogen Antibodies

Product name

AKT Pan Monoclonal Antibody (E.32.10)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody. This antibody is not cross-reactive with other related proteins.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

E.32.10

Vial size

100 µL

Concentration

19 µg/mL

Storage

-20°C

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Akt (pan) in HeLa cells, insulin-treated, using an Akt (pan) monoclonal antibody (Product # MA5-14999) showing translocation of Akt from the cytoplasm to the membrane upon insulin treatment. Actin filaments have been labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Akt (pan) in HeLa cells, serum-starved, using an Akt (pan) monoclonal antibody (Product # MA5-14999) showing translocation of Akt from the cytoplasm to the membrane upon insulin treatment. Actin filaments have been labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of Akt (pan) in paraffin-embedded human melanoma using an Akt (pan) monoclonal antibody (Product # MA5-14999).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of Akt (pan) in untreated Jurkat cells using a Akt (pan) monoclonal antibody (Product # MA5-14999) (blue) compared to a nonspecific negative control antibody (red).

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of Akt was performed using cross-linked chromatin from 1 x 106 HCT116 colon carcinoma cells treated with serum for 0, 15, 30, and 60 minutes. Immunoprecipitation was performed using a multiplex microplate Matrix ChIP assay (see reference for Matrix ChIP protocol: http://www.ncbi.nlm.nih.gov/pubmed/22098709) with 1.0 µL/100 µL well volume of an Akt monoclonal antibody (Product # MA5-14999). Chromatin aliquots from ~1 x 105 cells were used per ChIP pull-down. Quantitative PCR data were done in quadruplicate using 1 µL of eluted DNA in 2 µL SYBR real-time PCR reactions containing primers to amplify -15kb upstream of the Egr1 gene or exon-1 of Egr1. PCR calibration curves were generated for each primer pair from a dilution series of sheared total genomic DNA. Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the Egr-1 locus is shown above the data where boxes represent exons (black boxes = translated regions, white boxes = untranslated regions); the zigzag line represents an intron; and the straight line represents upstream sequence. Regions amplified by Egr-1 primers are represented by black bars. Data courtesy of the Innovators Program.