EAR1 Rabbit pAb

Blank control (Black line): Molt4 (Black).
Primary Antibody (green line): Rabbit Anti-EAR1 antibody (AP94717)
Dilution: 1 µg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF647
Dilution: 1 µg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Sample:
Lane 1: Mouse Spleen tissue lysates
Lane 2: Mouse Bone tissue lysates
Primary: Anti-EAR1 (AP94717) at 1/200 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 17 kDa
Paraformaldehyde-fixed, paraffin embedded (rat pancreas tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (EAR1) Polyclonal Antibody, Unconjugated (AP94717) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.

Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC-P, IHC-F, IF
Reactivity	Rat, Human, Mouse
Host	Rabbit
Clonality	Polyclonal
Physical State	Liquid
Immunogen	KLH conjugated synthetic peptide derived from mouse Eosinophil cationic protein 1
Epitope Specificity	65-155/155
Isotype	IgG
Purity	affinity purified by Protein A
Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
SUBCELLULAR LOCATION	Secreted. Note=Located in the matrix of eosinophil large specific granule, which are released following activation by an immune stimulus.
Important Note	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
Background Descriptions	Eosinophil derived neurotoxin (EDN) is a protein belonging to the ribonuclease (RNase) A superfamily. It has recently been found to have antiviral activity against respiratory syncytial virus and human immunodeficiency virus in vitro.

Additional Information
Dilution	WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1ug/Test
Storage	Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

For Research Use Only. Not For Use In Diagnostic Procedures.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.