Invitrogen

Calpastatin Polyclonal Antibody

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Calpastatin Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of Calpastatin on paraffin-embedded human intestinal cancer tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with Calpastatin polyclonal antibody (Product# PA5-78925) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Calpastatin Antibody in Western Blot (WB) — Western blot was performed using Anti-Calpastatin Polyclonal Antibody (Product # PA5-78925) and bands at 80-130 kDa corresponding to Calpastatin was observed in all the cell lines tested. Whole cell extracts (30 µg lysate) of A549 (Lane 1) and HeLa (Lane 2) were electrophoresed using Novex® NuPAGE® 4-12% % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Product Details

PA5-78925

Applications	Tested Dilution	Publications
Western Blot (WB)	0.1-0.5 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	0.5-1 µg/mL	-
Immunohistochemistry (Frozen) (IHC (F))	0.5-1 µg/mL	-
Immunocytochemistry (ICC/IF)	0.5-1 µg/mL	-
Flow Cytometry (Flow)	1-3 µg/1x10^6 cells	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A synthetic peptide corresponding to a sequence in the middle region of human Calpastatin.
Conjugate	Unconjugated
Form	Lyophilized
Concentration	500 µg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS with 5mg BSA
Contains	0.05mg sodium azide
Storage conditions	-20°C
RRID	AB_2746041

Product Specific Information

The synthetic peptide sequence is 275-310aa, QEKKRKVEKDTMSDQALEALSASLGTRQAEPELDLR

Add 0.2 mL of distilled water, will yield a concentration of 500 µg/mL.

Target Information

Calpastatin is an endogenous calpain (calcium-dependent cysteine protease) inhibitor. It consists of an N-terminal domain L and four repetitive calpain-inhibition domains (domains 1-4), and it is involved in the proteolysis of amyloid precursor protein. The calpain/calpastatin system is involved in numerous membrane fusion events, such as neural vesicle exocytosis and platelet and red-cell aggregation. The protein is also thought to affect the expression levels of genes encoding structural or regulatory proteins.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.