Recommended Dilution:Western Blot Analysis: 1ug/ml detects AIF from a mouse liver preparation.
Mouse liver protein preparation was resolved by electrophoresis, transferred to nitrocellulose and probed with AIF (1ug/ml).
Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system
Optimal dilutions to be determined by the researcher.
Western Blot Protocol 1.
Perform SDS-PAGE on a sample and transfer the proteins to nitrocellulose.
Wash the blotted nitrocellulose twice with water.
2.
Block the blotted nitrocellulose in freshly prepared TBS containing 3% nonfat dry milk with 0.1% Tween-20 (TBST-MLK) for 1 hour at RT with constant agitation.
3.
Incubate the nitrocellulose with 1ug/ml of AIF, A1059-77 diluted in freshly prepared TBS-MLK for three hours with agitation at RT4.
Wash the nitrocellulose twice with water.
5.
Incubate the nitrocellulose in the secondary reagent of choice (1:3000 dilution was used) in TBS-MLK overnight at 4ºC with agitation.
6.
Wash the nitrocellulose with water three times.
7.
Wash the nitrocellulose in TBS-0.1% Tween 20 for 30 minutes with agitation at RT.
8.
Rinse the nitrocellulose in water for 30 minutes with agitation at RT.
9.
Use detection method of choice (enhanced chemiluminescence was used).
Storage and Stability:Store product at 4°C in the dark.
DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid.
Dilute required amount only prior to immediate use.
Further dilutions can be made in assay buffer.
Caution: PE conjugates are sensitive to light.
For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Note: Applications are based on unconjugated antibody.
仕様
Size：100ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by Protein A affinity chromatography.
Form：Supplied as a liquid in PBS, pH 7.2. No preservative added. Labeled with R-Phycoerythrin (PE).
Specificity：Recognizes AIF, Mr 57kD. Another non-specific protein was also detected at Mr 120kD. Species Crossreactivity: Mouse. Predicted for human and rat based on sequence homology.
Isotype：IgG
Calc Applications Abbrev：WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：KLH-conjugated, synthetic peptide corresponding to amino acids 593–606 of human Apoptosis Inducing Factor, AIF (C-KDGEQHEDLNEV-AK), containing a N-terminal cysteine. The immunizing sequence is identical in mouse and rat.
