Secretases are proteolytic enzymes involved in the processing of amyloid beta (Aβ). The β/γ Secretase Antibody Sampler Kit includes essential components of the secretase proteolytic machinery including BACE1, nicastrin, PSEN1, and PSEN2. The β/γ secretase proteins are often found associated with cellular membranes such as the endosome and the plasma membrane. These antibodies are therefore ideal for detection of protein expression levels and localization within cellular compartments. The tubulin β3 antibody is included as a loading control for WB or visualization of neurons by immunostaining.

 

β/γ Secretase Antibody Sampler Kit contents:

Specificity

Clone

Format

Reactivity

Application

BACE1

A17035K

Purified

Hu

WB, Direct ELISA

Nicastrin

9C3

Purified

Hu, Ms, Rat

IHC-P, WB

Presenilin 1 (N terminus)

NT1

Purified

Hu

WB

Presenilin 2

PS2

Purified

Hu

WB

Tubulin β 3 (TUBB3)

TUJ1

Purified

Hu, Ms, Rat

IHC-P, ICC, WB

Clone A17035K recognizes beta-secretase, also known as beta-site APP cleavage enzyme 1 (BACE1), a protease expressed in neurons. The cleavage of APP at the N-terminus of the Aβ domain by BACE1 releases a soluble extracellular fragment called sAPPβ. The remaining portion of APP, which is membrane-bound, is termed C-terminal fragment (C99).

 

 

 

 

Though not catalytically active itself, Nicastrin is a critical component of gamma-secretase, and is responsible for promoting maturation and localization of other units to the enzymatic complex. Clone 9C3 can detect Nicastrin by western blotting and IHC.

 

 

 

 

Clone NT1 can be used to detect presenilin 1, another component of gamma-secretase. Gamma-secretase is responsible for the processing of the C99 fragment to release Aβ isoforms, including Aβ 1-40 and Aβ 1-42, and a fragment known as the APP intracellular domain (AICD). Increased Aβ production has been partly attributed to mutations in the APP or PSEN genes. These mutations favor the processing of APP by β- and γ-secretases leading to elevated levels of Aβ.

 

 

 

Clone PS2 can be used to detect presenilin 2, which is also a component of gamma-secretase, by western blotting. Mutations in presenilin 1 and 2 are major causative factors of early onset familial Alzheimer’s disease.

 

 

 

 

Clone TUJ1 is highly reactive to class III β-tubulin, a cytoskeletal protein expressed in neurons. Clone TUJ1 does not react with β-tubulin found in glial cells. Immunostaining with TUJ1 allows visualization of cell bodies, dendrites, and axons.

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