ENCAB000AIT
Antibody against Homo sapiens MXI1, Mus musculus MXI1
Homo sapiens
K562
characterized to standards
Homo sapiens
HeLa-S3, GM12878, HepG2, neural cell
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
Mus musculus
any cell type or tissue
partially characterized
- Status
- released
- Source (vendor)
- RD Systems
- Product ID
- AF4185
- Lot ID
- ZIJ0107031
- Characterized targets
- MXI1 (Homo sapiens), MXI1 (Mus musculus)
- Host
- goat
- Clonality
- polyclonal
- Purification
- affinity
- Isotype
- IgG
- Antigen description
- Raised against E. coli-derived rhMxi1.
- External resources
Characterizations
MXI1 (Homo sapiens)
neural cell
exempt from standards
- Caption
- The ENCODE antibody standards document exempts some valuable and/or limiting samples from primary characterizations for well-characterized antibodies. They are given exemptions so that the samples can be conserved to carry out the downstream experiments.
- Submitter comment
- We do not have any of the H1-derived neurons that were distributed to the consortium for analysis in ENCODE2 left.
- Reviewer comment
- This cell type was exempted from primary characterization of antibody ENCAB000AIT by the ENCODE antibody review panel on October 17, 2016
- Submitted by
- Jessika Adrian
- Lab
- Michael Snyder, Stanford
- Grant
- U54HG006996
- Download
- H1neuron_primary.png
MXI1 (Homo sapiens)
not reviewed
- Submitted by
- Michael Snyder
- Lab
- Michael Snyder, Stanford
- Grant
- U54HG004558
- Download
- human_MXI1_validation_Snyder.pdf
MXI1 (Homo sapiens)
not reviewed
- Submitted by
- Michael Snyder
- Lab
- Michael Snyder, Stanford
- Grant
- U54HG004558
- Download
- human_MXI1_validation_Snyder.pdf
MXI1 (Homo sapiens)
K562
compliant
- Caption
- Immunoprecipitation of MXI from K562 cells using AF4185. Lane 1: input nuclear lysate, Lane 2: material immunoprecipitated with ab87525, Lane 3: material immunoprecipitated using control IgG. Band A was excised from the gel and subject to analysis by mass spectrometry. Expected size: 26kD
- Submitted by
- Kathrina Onate
- Lab
- Michael Snyder, Stanford
- Grant
- U54HG004558
- Download
- IPMS_MXI1_Snyder_AIT.png
MXI1 (Mus musculus)
not reviewed
- Submitted by
- Michael Snyder
- Lab
- Michael Snyder, Stanford
- Grant
- RC2HG005602
- Download
- mouse_Mxi1_validation_Snyder.pdf
MXI1 (Mus musculus)
not reviewed
- Submitted by
- Michael Snyder
- Lab
- Michael Snyder, Stanford
- Grant
- RC2HG005602
- Download
- mouse_Mxi1_validation_Snyder.pdf
MXI1 (Homo sapiens)
compliant
- Caption
- IP followed by mass spectrometry: Briefly, protein was immunoprecipitated from K562 whole cell lysates using AF4185, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGE Bis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was silver-stained, gel fragments corresponding to the bands indicated were excised and destained using the SilverSNAP Stain for Mass Spectrometry (Pierce). Then proteins were trysinized using the in-gel digestion method. Digested proteins were analyzed on an LTQ-Orbitrap (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Protein false discovery rate < 1%, 2 peptides per protein minimum).
- Submitted by
- Kathrina Onate
- Lab
- Michael Snyder, Stanford
- Grant
- U54HG004558
- Download
- Mxi1_final Sheet1.pdf
MXI1 (Homo sapiens)
HeLa-S3K562GM12878HepG2
exempt from standards
- Caption
- Western blot analysis of nuclear lysates prepared from multiple cells lines loaded in the order: HeLa-S3,K562,GM12878,HepG2 using the antibody AF4185. Molecular Weight: 26.062.
- Submitter comment
- We observed one major band of ~40kD in all cell lines assayed, along with bands of lesser intensity at ~50kD and ~32kD. The major band migrates somewhat slower than predicted for MXI1 (~26kD), but a similar sized band in the K562 IP was confirmed to contain MXI1 by mass spectrometry.
- Reviewer comment
- The major band detected is ~40kD and greater than the allowed 20% deviation from the expected size (~26kD) according to the standards but the secondary IP followed by mass spectrometry characterization in K562 showed that MXI1 is detected in the slower migrating band at that higher size, and likely the case for other cell lines as well.
- Submitted by
- Nathaniel Watson
- Lab
- Michael Snyder, Stanford
- Grant
- U54HG006996
- Download
- WB Mxi1 (AF4185).jpg