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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [1]
- Immunohistochemistry [1]
- Other assay [1]
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- Product number
- PA5-54271 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NEK1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.7 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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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.
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).
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.
