1. Antibodies
  2. Homo sapiens
  3. NFXL1

ENCAB208FGX

Antibody against Homo sapiens NFXL1

Homo sapiens
K562
characterized to standards
Homo sapiens
GM12878, MCF-7
characterized to standards with exemption
Homo sapiens
HepG2
partially characterized
Status
released
Source (vendor)
Bethyl Labs
Product ID
A303-197A
Lot ID
1
Characterized targets
NFXL1 (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Aliases
michael-snyder:AS-1020
External resources

Characterizations

NFXL1 (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not submitted for review by lab
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: HepG2 using the antibody A303-197A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 101.339.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
Download
 2.jpg
NFXL1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562, using the antibody A303-197A. The blot shows western blot analysis of input, flowthrough, immunoprecipitate and mock immunoprecipitate using IgG.
Reviewer comment
Not within 20% of expected value and not 50% of total signal.
Submitted by
Denis Salins
Lab
Michael Snyder, Stanford
Download
 4.jpg
NFXL1 (Homo sapiens)
GM12878
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: GM12878 using the antibody A303-197A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 101.339.
Submitter comment
Except target band, there is a stronger band between 50KD and 75KD. K562 Mass Spec proved that NFXL1 was also detected as rank 1 in this band.
Reviewer comment
Marked band not >50% of total immunoreactive signal in the lane but mass spec in K562 also detected the target factor at ~50kD signal as the top ranked hit.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NFXL1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line K562 using the antibody A303-197A. Lane 1: input nuclear lysate. Lane 2: material immunoprecipitated with antibody. Lane 3: material immunoprecipitated using control IgG. Marked bands were excised from gel and subjected to analysis by mass spectrometry. Target molecular weight: 101.339.
Reviewer comment
Runs smaller than expected size but NFXL1 is the top TF detected in mass spec in both analyzed immunoreactive bands.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NFXL1 (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
compliant
Caption
IP followed by mass spectrometry. Briefly, protein was immunoprecipitated from K562 nuclear cell lysates using the antibody A303-197A, and the IP fraction was loaded on a 10% polyacrylamide gel (NuPAGEBis-Tris Gel) and separated with an Invitrogen NuPAGE electrophoresis system. The gel was stained by ColloidialCoomassie G-250 stain, gel fragments corresponding to the bands indicated were excised. Then proteins were trypsinized using the in-gel digestion method. Digested proteins were analyzed on an Orbitrap Elite mass spectrometer (Thermo Scientific) by the nanoLC-ESI-MS/MS technique. Peptides were identified by the SEQUEST algorithm and filtered with a high confidence threshold (Peptide false discovery rate < 1%, 2 unique peptides per protein minimum, mass error < 10 ppm).
Submitter comment
RBM10 is RNA-binding protein.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NFXL1 (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: K562 using the antibody A303-197A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 101.339.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford
NFXL1 (Homo sapiens)
MCF-7
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Immunoprecipitation was performed on nuclear extracts from the cell line: MCF-7 using the antibody A303-197A. The image shows western blot analysis of input, flowthrough, immunoprecipitate, and mock immunoprecipitate using IgG. Target molecular weight: 101.339.
Submitter comment
1) same pattern as K562 IP. 2) Mass Spec result indicates that the both bands(~65-100kd) are NFXL1
Reviewer comment
Multiple immunoreactive bands seen but the banding pattern is consistent with the IP in K562 that was mass-spec'd and TF was detected in both marked bands.
Submitted by
Nathaniel Watson
Lab
Michael Snyder, Stanford