Anti-human APPL1 Antibody, Rabbit pAb

APPL1, an adaptor protein containing an NH2-terminal Bin/Amphiphiphysin/Rvs (BAR) domain, a central pleckstrin homology (PH) domain and a COOH-terminal phosphotyrosine binding (PTB) domain [1] was originally identified as an interacting partner of Akt in a yeast two-hybrid assay using Akt2 as a bait [2]. APPL1 binds to a number of cell surface receptors (TrkA [3,4], DCC [5], adiponectin [6,7], FSH [8]) and intracellular signaling molecules (small GTPase Rab5 [9], GIPC [4] and inositol 5-phosphatase [10]), suggesting that APPL1 may act as a common relay to coordinate diverse signaling cascades. APPL1 potentiates insulin-mediated Akt activation by counteracting the effect of the Akt inhibitor TRB3 [11].

Recombinant full-length human APPL1 expressed in E.coli.

The antibody detects several types of APPL1 in different species such as human, monkey, mouse, rat etc. (about 85kDa).

Liquid in phosphate-buffered saline (PBS).

Store at –20°C for less than one week. For long-term storage, aliquot and freeze at -70°C. Avoid repeated freeze/defrost cycles.

Western blot - This antibody can be used at 0.1-0.2 μg/mL with the appropriate secondary reagents to detect APPL1.

ELISA - This antibody can be used at 2.0-5.0 μg/mL with the appropriate secondary reagents to detect APPL1.

Immunoprecipitation – See reference [6], [11].

Immunostaining - This antibody can be used at 1.0-2.0 μg/mL with the appropriate secondary reagents to detect APPL1.

Antigen affinity-purified

1. Hosch, S.E., J.M. Olefsky, and J.J. Kim, APPLied mechanics: uncovering how adiponectin modulates insulin action. Cell Metab, 2006. 4(1): p. 5-6.

2. Mitsuuchi, Y., et al., Identification of a chromosome 3p14.3-21.1 gene, APPL, encoding an adaptor molecule that interacts with the oncoprotein-serine/threonine kinase AKT2. Oncogene., 1999. 18(35): p. 4891-8.

3. Lin, D.C., et al., APPL1 associates with TrkA and GIPC1, and is required for NGFmediated signal transduction. Mol Cell Biol, 2006. 25: p. 25.

4. Varsano, T., et al., GIPC is recruited by APPL to peripheral TrkA endosomes and regulates TrkA trafficking and signaling. Mol Cell Biol, 2006. 26(23): p. 8942-52.

5. Liu, J., et al., Mediation of the DCC apoptotic signal by DIP13 alpha. J Biol Chem., 2002. 277(29): p. 26281-5. Epub 2002 May 14.

6. Cheng, K.K., et al., Adiponectin-induced endothelial nitric oxide synthase activation and nitric oxide production are mediated by APPL1 in endothelial cells. Diabetes, 2007. 56(5): p. 1387-94.

7. Mao, X., et al., APPL1 binds to adiponectin receptors and mediates adiponectin signalling and function. Nat Cell Biol., 2006. 8(5): p. 516-23. Epub 2006 Apr 16.

8. Nechamen, C.A., et al., Human follicle-stimulating hormone (FSH) receptor interacts with the adaptor protein APPL1 in HEK 293 cells: potential involvement of the PI3K pathway in FSH signaling. Biol Reprod., 2004. 71(2): p. 629-36. Epub 2004 Apr 7.

9. Miaczynska, M., et al., APPL proteins link Rab5 to nuclear signal transduction via an endosomal compartment. Cell., 2004. 116(3): p. 445-56.

10. Erdmann, K.S., et al., A role of the Lowe syndrome protein OCRL in early steps of the endocytic pathway. Dev Cell, 2007. 13(3): p. 377-90.

11. Cheng, K.K., et al., APPL1 potentiates insulin-mediated inhibition of hepatic glucose production and alleviates diabetes via Akt activation in mice. Cell Metab, 2009. 9(5): p. 417-27.

1. Cheng KK, Zhu W, Chen B, Wang Y, Wu D, Sweeney G, Wang B, Lam KS, Xu A. The adaptor protein APPL2 inhibits insulin-stimulated glucose uptake by interacting with TBC1D1 in skeletal muscle. Diabetes. 2014 Nov 1;63(11):3748-58.
2. Yau SY, Li A, Hoo RL, Ching YP, Christie BR, Lee TM, Xu A, So KF. Physical exercise-induced hippocampal neurogenesis and antidepressant effects are mediated by the adipocyte hormone adiponectin. Proceedings of the National Academy of Sciences. 2014 Nov 4;111(44):15810-5.
3. Dadson K, Kovacevic V, Rengasamy P, Kim GH, Boo S, Li RK, George I, Schulze PC, Hinz B, Sweeney G. Cellular, structural and functional cardiac remodelling following pressure overload and unloading. International journal of cardiology. 2016 Aug 1;216:32-42.
4. Jiang X, Zhou Y, Wu KK, Chen Z, Xu A, Cheng KK. APPL1 prevents pancreatic beta cell death and inflammation by dampening NFκB activation in a mouse model of type 1 diabetes. Diabetologia. 2017 Mar 1;60(3):464-74.
5. Fan L, Ye H, Wan Y, Qin L, Zhu L, Su J, Zhu X, Zhang L, Miao Q, Zhang Q, Zhang Z. Adaptor protein APPL1 coordinates HDAC3 to modulate brown adipose tissue thermogenesis in mice. Metabolism. 2019 Nov 1;100:153955.
6. Botta A, Liu Y, Wannaiampikul S, Tungtrongchitr R, Dadson K, Park TS, Sweeney G. An adiponectin-S1P axis protects against lipid induced insulin resistance and cardiomyocyte cell death via reduction of oxidative stress. Nutrition & metabolism. 2019 Dec;16(1):1-2.
7. Gao C, Yan T, Chen X, Cheng KK, Xu A, Shen J. APPL2 Negatively Regulates Olfactory Functions by Switching Fate Commitments of Neural Stem Cells in Adult Olfactory Bulb via Interaction with Notch1 Signaling. Neuroscience Bulletin. 2020 Sep;36:997-1008.