ENCAB686SDS

Homo sapiens

K562

characterized to standards

Method: knockdown or knockout

compliant

Caption: Western blot following CRISPR against RYBP in HepG2 whole cell lysate using RYBP specific antibody. Lane 1 is a ladder, lane 2 is HepG2 non-targeting control knockdown, lane 3 and 4 are two different CRISPR against RYBP. RYBP protein appears as the green arrow, Beta-actin serves as a control and appears in red arrow.
Submitted by: Xintao Wei
Lab: Brenton Graveley, UConn
Grant: U41HG009889
Download: RYBP-HEPG2-CRISPR-PA5-19943.png
Documents: encode:TF_Antibody_Characterization_ENCODE3_May2016.pdf

K562

Method: immunoprecipitation

compliant

Caption: IP-WB analysis of K562 whole cell lysate using the RYBP specific antibody, PA5-19943. Lanes 1 and 2 are 2.5% of five million whole cell lysate input and 50% of IP enrichment, respectively, using a normal IgG antibody. Lane 3 is 50% of IP enrichment from five million whole cell lysate using the RYBP-specific antibody, PA5-19943. The same antibody was used to detect protein levels via Western blot. This antibody passes preliminary validation and will be further pursued for secondary validation. *NOTE* Protein sizes are taken from Genecards.org and are only estimates based on sequence. Actual protein size may differ based on protein characteristics and electrophoresis method used.
Submitter comment: Immunoprecipitation lane (3) exhibits a large deviation from the expected protein size, which we believe is due to performing the characterization using the Jess Western Blotting system. This system has a documented tendency for some proteins to run differently than with more traditional methods due to their compositions and interactions with the electrophoretic components. The image also closely resembles the example image provided by the vendor.
Submitted by: Steven Blue
Lab: Gene Yeo, UCSD
Grant: U54HG007005
Download: ThermoFisher_PA5-19943_UF2796583_RYBP.png
Documents: encode:RBP_Antibody_Characterization_ENCODE3_Nov2016.pdf