1. Antibodies
  2. Homo sapiens
  3. DROSHA

ENCAB936LAQ

Antibody against Homo sapiens DROSHA

Homo sapiens
K562
characterized to standards
Homo sapiens
HepG2
characterized to standards with exemption
Homo sapiens
any cell type or tissue
partially characterized
Status
released
Source (vendor)
Bethyl Labs
Product ID
A301-886A
Lot ID
1
Characterized targets
DROSHA (Homo sapiens)
Host
rabbit
Clonality
polyclonal
Purification
affinity
Isotype
IgG
External resources

Characterizations

DROSHA (Homo sapiens)
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not submitted for review by lab
Caption
IP-WB analysis of K562 whole cell lysate using DROSHA specific antibody. Lane 1 is 2.5% of 0.5mg input lysate, lane 2 is 2.5% of supernatant after immunoprecipitation and Lane 3 is 50% of IP enrichment using rabbit polyclonal Drosha Antibody. This antibody passes preliminary validation and will be further pursued for primary and secondary validation. Secondary validation is necessary because the predicted band in not recognized in the 'input/supernatant' sample due to low expression level.
Submitted by
Balaji Sundararaman
Lab
Gene Yeo, UCSD
DROSHA (Homo sapiens)
Method: immunoprecipitation followed by mass spectrometry
Attachment PDF Icon
not reviewed
Caption
IP followed by mass spectrometry. Protein was immunoprecipitated from HepG2 whole cell lysates using the antibody A301-886A, loaded on a 4-12% NuPAGE Bis-Tris gel, and separated via electrophoresis. Using a reference western blot done in parallel, gel pieces corresponding to the sections indicated were excised and submitted for analysis by the UCSD Biomolecular and Proteomics Mass Spectrometry Facility.
Reviewer comment
Not all immunoreactive bands were excised for mass spec analysis but seeing as there is already a compliant secondary characterization for this antibody, review of this mass spec is not required.
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
DROSHA (Homo sapiens)
Method: knockdown or knockout
Attachment from submitter
Attachment from submitters
compliant
Caption
Western blot following shRNA against DROSHA in K562 whole cell lysate using DROSHA specific antibody. Lane 1 is a ladder, lane 2 is K562 non-targeting control knockdown, lane 3 and 4 are two different shRNAs against DROSHA.DROSHA protein appears as the green band, Tubulin serves as a control and appears in red.
Submitted by
Xintao Wei
Lab
Brenton Graveley, UConn
DROSHA (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
exempt from standards
Caption
Representative image of immunoprecipitation performed on whole cell extracts from the HepG2 cell line using the DROSHA-specific antibody A301-886A. Lane 1: Input from IP using control IgG. Lane 2: Immunoprecipitated material using control IgG. Lane 3: Input from IP using DROSHA antibody. Lane 4: Immunoprecipitated material using DROSHA antibody. Outlined regions were excised from gel and subjected to analysis by mass spectrometry. Target molecular weight: 159.32 kDa.
Submitter comment
We confirmed by mass spec that the factor of interest can be detected in the marked band.
Reviewer comment
Originally deemed not compliant by another reviewer on the basis of the marked band not comprising of at least 50% of the total signal in the lane. However, given that it is difficult to judge quantitatively and is fairly close to 50% (+/- some margin) and shown by mass spec to contain the factor of interest, we feel that it should be exempt from the hard cutoff.
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
DROSHA (Homo sapiens)
HepG2
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
not compliant
Caption
IP-Western Blot analysis of HepG2 whole cell lysate using DROSHA specific antibody. Lane 1 is 1% of twenty million whole cell lysate input and lane 2 is 25% of IP enrichment using rabbit normal IgG (lanes under 'IgG'). Lane 3 is 1% of twenty million whole cell lysate input and lane 4 is 10% IP enrichment using rabbit polyclonal anti-DROSHA antibody (lanes under 'DROSHA').
Submitter comment
band at 159kDa is not 50% of lane signal
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD
DROSHA (Homo sapiens)
K562
Method: immunoprecipitation
Attachment from submitter
Attachment from submitters
compliant
Caption
IP-Western Blot analysis of K562 whole cell lysate using DROSHA specific antibody. Lane 1 is 1% of twenty million whole cell lysate input and lane 2 is 10% of IP enrichment using rabbit normal IgG (lanes under 'IgG'). Lane 3 is 1% of twenty million whole cell lysate input and lane 4 is 10% IP enrichment using rabbit polyclonal anti-DROSHA antibody (lanes under 'DROSHA').
Submitted by
Steven Blue
Lab
Gene Yeo, UCSD