PA5-54271

antibody from Invitrogen Antibodies

Targeting: NEK1 KIAA1901, NY-REN-55

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-54271

Provider

Invitrogen Antibodies

Product name

NEK1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant full-length protein

Description

Immunogen sequence: NLKAQEDEKG KQNLSDTFEI NVHEDAKEHE KEKSVSSDRK KWEAGGQLVI PLDELTLDTS FSTTERHTVG EVIKLGPNGS PRRAWGKSPT DSVLKILGEA ELQLQTELLE NTTIRSEISP EGEKYKPLIT GEKKVQCISH EINPSA

Concentration

0.7 mg/mL

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of Nek1 was performed by loading 100 µg of WT (lane 1) and NEK1 CRISPR KO (lane 2) HeLa cell lysates in RIPA buffer onto a 5-16% gradient polyacrylamide gel. Proteins on the blots were visualized with Ponceau staining (below immunoblot). Proteins were transferred to nitrocellulose membrane and blocked in 5% milk for 1 hr. Nek1 was detected at approximately 143 kDa (designated by the black arrow) using a Nek1 polyclonal antibody (Product # PA5-54271) at a dilution of 1:1000 in 5% BSA in TBS with 0.1% Tween 20 (TBST) overnight at 4 deg. The peroxidase-conjugated secondary antibody (Product # 65-6120) was diluted to 0.2 µg/mL in TBST with 5% milk for 1 hr. Chemiluminescent detection was performed using Pierce ECL Western Blotting Substrate (Product # 32106). Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical staining of NEK1 in human stomach tissue shows strong cytoplasmic positivity in glandular cells. Samples were probed using a NEK1 Polyclonal Antibody (Product # PA5-54271).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of NEK1 was performed on HeLa cell lysates. Antibody-bead conjugates were prepared by adding 1 µg of NEK1 polyclonal antibody (Product # PA5-54271) with 30 µL of protein A -Sepharose beads and rocked overnight at 4 deg. 1 µg of lysate was incubated with antibody-bead conjugate for 2 hrs at 4 deg. After multiple washes, 10% starting material (SM), 10% unbound fraction (UB) and immunoprecipitated fraction (IP) were processed for immunoblot using a different NEK1 polyclonal antibody (Product # PA5-78074). Ponceau stained transfer of blot is shown. Data courtesy of YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies using knockout validation.