700632

antibody from Invitrogen Antibodies

Targeting: PTK2B CADTK, CAKB, FAK2, PTK, PYK2, RAFTK

Western blot (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

700632

Provider

Invitrogen Antibodies

Product name

Phospho-PYK2 (Tyr402) Recombinant Rabbit Monoclonal Antibody (17H4L19)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.

Antibody clone number

17H4L19

Concentration

0.5 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-FAK2 pTyr402 in whole cell extract (60 µg per lane) from A431 cells (lane 1) and Raji cells (lane 2) using a Phospho-FAK2 pTyr402 recombinant rabbit monoclonal antibody (Product # 700632) at a dilution of 5 µg/mL. Detection was performed using an HRP-conjugated goat anti-rabbit secondary antibody at a dilution of 1:5000 followed by chemiluminescence (ECL). Results show a band at ~115 kDa.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of Phospho-FAK2/PYK2 (pTyr402) performed using cross-linked chromatin from 1 x 10^6 HCT116 human colon carcinoma cells treated with serum for 0, 15, 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 a Phospho-FAK2/PYK2 (pTyr402) rabbit monoclonal antibody (Product # 700632). Chromatin aliquots from ~1 x 10^5 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 human Egr-1 locus, or exon-1 or exon-2 of Egr-1. 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.