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
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
- Grant
- U54HG006996
- Download
- 2.jpg
NFXL1 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- 4.jpg
NFXL1 (Homo sapiens)
GM12878
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
- Grant
- U54HG006996
NFXL1 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- MS1038_1_NFXL1-A303-197A-3.jpg
NFXL1 (Homo sapiens)
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
- Grant
- U54HG006996
- Download
- NFXL1_A303-197A final.pdf
NFXL1 (Homo sapiens)
K562
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
- Grant
- U54HG006996
- Download
- NFXL1_A303-197A.jpg
NFXL1 (Homo sapiens)
MCF-7
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
- Grant
- U54HG006996
- Download
- Scan_20170420 (3).jpg