700028

antibody from Invitrogen Antibodies

Targeting: JAK1 JAK1A, JAK1B, JTK3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

700028

Provider

Invitrogen Antibodies

Product name

Phospho-JAK1 (Tyr1022, Tyr1023) Recombinant Rabbit Monoclonal Antibody (59H4L5)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with mouse, rat, chimpanzee, Rhesus monkey, bovine, canine, porcine, equine, Xenopus and zebrafish based on sequence homology.

Antibody clone number

59H4L5

Concentration

0.5 mg/mL

References

Submitted references

Recombinant human erythropoietin and interferon-β-1b protect against 3-nitropropionic acid-induced neurotoxicity in rats: possible role of JAK/STAT signaling pathway.

Sayed RH, Ghazy AH, Yammany MFE
Inflammopharmacology 2022 Apr;30(2):667-681

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Phospho-JAK1 pTyr1022/1023 in untreated 3T3-L1 lysates (lane 2) or 25 ng/mL LIF-stimulated 3T3-L1 lysates (lane 1) using a Phospho-JAK1 pTyr1022/1023 recombinant rabbit monoclonal antibody (Product # 700028) at a dilution of 1 µg/mL.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of JAK1 (pY1022/1023) was done on 70% confluent log phase COLO 205 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with JAK1 (pY1022/1023) Recombinant Rabbit Monoclonal Antibody (Product # 700028) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing membrane localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin immunoprecipitation analysis of Phospho-JAK1 (pTyr1022+1023) was 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-JAK1 (pTyr1022+1023) rabbit monoclonal antibody (Product # 700028). 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 the promoter region of human UBE2B, or the imprinting control region (ICR) of the human H19 locus. 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 human UBE2B and H19 loci are shown above the data where boxes represent exons (grey boxes = translated regions, white boxes = untranslated regions), the zigzag lines represent introns, and the straight line represents upstream sequence. Regions amplified by UBE2B and H19 primers are represented by black bars. Data courtesy of the Innovators Program.