702779

antibody from Invitrogen Antibodies

Targeting: SACS ARSACS, DKFZp686B15167, DNAJC29, KIAA0730, PPP1R138, SPAX6

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

702779

Provider

Invitrogen Antibodies

Product name

Anti-Sacsin DNAJ domain Antibody (4H18L14), ABfinity™ Rabbit Monoclonal

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with Monkey, Rabbit, Cat

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

4H18L14

Vial size

100 µg

Concentration

0.5 mg/ml

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on different concentrations of recombinant Sacsin DNAJ domain protein. The blots were probed with ABfinity™ Anti-Sacsin DNAJ domain Recombinant Rabbit Monoclonal Antibody (Product # 702779, 2.5 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/ml, 1:4000 dilution). A 35 kDa band corresponding to Sacsin DNAJ domain was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, SH-SY5Y cells were fixed and permeabilized for detection of endogenous Sacsin DNAJ domain using ABfinity™ Anti- Sacsin DNAJ domain Recombinant Rabbit Monoclonal Antibody (Product# 702779, 5 µg/ml) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Sacsin DNAJ domain (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents MitoTracker® Red CMXRos (Product # M7512, 50nM). Panel d) is a composite image of Panels a, b and c clearly demonstrating partial mitochondrial overlap of Sacsin DNAJ domain. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.