- 33-3700 - Invitrogen Antibodies ANK2 antibody

Submitted references PERM1 interacts with the MICOS-MIB complex to connect the mitochondria and sarcolemma via ankyrin B.

Bock T, Türk C, Aravamudhan S, Keufgens L, Bloch W, Rozsivalova DH, Romanello V, Nogara L, Blaauw B, Trifunovic A, Braun T, Krüger M
Nature communications 2021 Aug 12;12(1):4900

Coupling of the Na+/K+-ATPase to Ankyrin B controls Na+/Ca2+ exchanger activity in cardiomyocytes.

Skogestad J, Aronsen JM, Tovsrud N, Wanichawan P, Hougen K, Stokke MK, Carlson CR, Sjaastad I, Sejersted OM, Swift F
Cardiovascular research 2020 Jan 1;116(1):78-90

Activation of glial FGFRs is essential in glial migration, proliferation, and survival and in glia-neuron signaling during olfactory system development.

Gibson NJ, Tolbert LP, Oland LA
PloS one 2012;7(4):e33828

Roles of specific membrane lipid domains in EGF receptor activation and cell adhesion molecule stabilization in a developing olfactory system.

Gibson NJ, Tolbert LP, Oland LA
PloS one 2009 Sep 29;4(9):e7222

Structural requirements for interaction of sodium channel beta 1 subunits with ankyrin.

Malhotra JD, Koopmann MC, Kazen-Gillespie KA, Fettman N, Hortsch M, Isom LL
The Journal of biological chemistry 2002 Jul 19;277(29):26681-8

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of rat brain homogenates using Ms anti-Ankyrin B (Product # 33-3700).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Ankyrin B was performed by loading 20 µg of Rat Heart (lane1) and SH-SK5Y (lane2) lysate using Novex®NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and proteins were transferred to a PVDF membrane with overnight wet transfer setup at 4°C and blocked with 5% skim milk for 1 hour at room temperature. Ankyrin B was detected at ~ 220 kDa using Ankyrin B Mouse Monoclonal Antibody (Product # 33-3700) at 0.5-1 µg/mL in 5% skim milk for 3 hour at room temperature on a rocking platform. Goat Anti-Mouse - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Ankyrin B was performed by loading 20 µg of Rat Heart (lane1) and SH-SK5Y (lane2) lysate using Novex®NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and proteins were transferred to a PVDF membrane with overnight wet transfer setup at 4°C and blocked with 5% skim milk for 1 hour at room temperature. Ankyrin B was detected at ~ 220 kDa using Ankyrin B Mouse Monoclonal Antibody (Product # 33-3700) at 0.5-1 µg/mL in 5% skim milk for 3 hour at room temperature on a rocking platform. Goat Anti-Mouse - HRP Secondary Antibody (Product # 62-6520) at 1:4000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Ankyrin B was done on 70% confluent log phase SHSY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Ankyrin B Mouse monoclonal Antibody (Product # 33-3700) at 1 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 PERM1 associates with ankyrin B and the interaction with the MICOS-MIB complex is dependent on the transmembrane helix. a PERM1 interacts with ANKB, as indicated by immunoprecipitation of FLAG-tagged Perm1 and HA-tagged ankyrin-B in TREx-293 cells stably expressing Perm1 -FLAG. Protein lysates were immunoprecipitated with anti-FLAG beads and anti-HA beads and immunoblotted with anti-FLAG and anti-HA antibodies. b Immunohistochemical staining of soleus muscle revealed co-localization of PERM1 (red) with ANKB (green). Scale, 50 um. c Mean fractional intensity of ANKB signals in bins 1-6 from Perm1 -/- and wild-type soleus muscle fibers. ANKB staining intensity was reduced at subsarcolemmal sites and increased within interfibrillar mitochondria in Perm1 -/- muscles compared to wild-type controls ( n = 4 mice per group, 2299 quantified fibers). Box plot (c) represents the median, 25th, and 75th percentiles, maximum and minimum are connected through whiskers. Outliers are defined as Q 1 -1.8 IQR and Q 3 +1.8 IQR. (c) unpaired two-sided Mann-Whitney U -test. Source data are provided as Source Data file.

33-3700

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