FHL1
Reactivity: Human
WB, IHC
Host: Rabbit
Monoclonal
unconjugated
Application Notes
WB (1:500 - 1:2000), IF/IC (1:50 - 1:100)
Restrictions
For Research Use only
Validation #101371
(Immunofluorescence)
by
Alexander Lab, Jacobs School of Medicine and Biomedical Sciences, Department of Medicine, University of Buffalo
No.
#101371
Date
07/19/2017
Antigen
FHL-1
Lot Number
AL1F04A
Method validated
Immunofluorescence
Positive Control
MRL/lpr lupus mice (14 weeks of age)
Negative Control
FH knockout MRL/lpr lupus mice
Notes
Passed. ABIN2966633 works well on murine kidney samples and shows the expected CFH1 localization.
Validation Images
IF staining with ABIN2966633 of kidneys harvested from 14 week old MRL/lpr mice sufficient and deficient in factor H. Cryosections were fixed, stained and images acquired as described in the protocol.
Full Methods
Primary Antibody
ABIN2966633
Secondary Antibody
goat anti-rabbit rhodamine conjugated antibody (ThermoFisher Scientific, T-2769, lot 6251)
Full Protocol
Prepare kidneys of wt and complement factor H ko MRL/lpr lupus mice of 14 weeks of age.
Snap-freeze kidneys in 2-methylbutane cooled on dry ice and keep at −80°C until use.
Sections were cut at 8µm using the cryostat.
Dry sections for 30min at RT.
Hydrate sections in PBS 2x for 15min at RT.
Fix cryostat sections with 4% paraformaldehyde (PFA) for 15min at RT.
Wash sections 2x with PBS for 15min at RT.
Incubate sections with primary rabbit anti-FHL-1 antibody (antibodies-online, ABIN2966633, AL1F04A) diluted 1:100 in PBS ON at 4°C.
Wash sections 2x for 15min with PBS.
Incubate sections with secondary goat anti-rabbit rhodamine conjugated antibody (ThermoFisher Scientific, T-2769, lot 6251) diluted 1:100 in PBS for 30min at RT.
Wash sections 2x for 15min with PBS.
Incubate sections with ActinGreen 488 ReadyProbes Reagent (ThermoFisher Scientific, R37122, lot 1700569) diluted 1:100 in PBS for 30min at RT.
Wash sections 2x for 15min with PBS.
Image acquisition on a Zeiss microscope (Carl Zeiss, Oberkochen, Germany).
Experimental Notes
Format
Liquid
Buffer
Liquid in 0.42 % Potassium phosphate, 0.87 % Sodium chloride, pH 7.3, 30 % glycerol, and 0.01 % sodium azide.
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C
Storage Comment
Shipped at 4°C. Upon delivery aliquot and store at -20°C for one year. Avoid freeze/thaw cycles.