Name: Anti-KCNJ5 (Kir3.4) Antibody
Brand: Alomone Labs
SKU: APC-027

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Overview

Cat #: APC-027

Alternative Name GIRK4, G protein-activated inward rectifier potassium channel 4, Cardiac inward rectifier, CIR, KATP-1

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: h, m, r

Immunogen

Peptide RNAMNQDMEIGVT(C), corresponding to amino acid residues 6-18 of rat K_ir3.4 (Accession P48548). Intracellular, N-terminus.

Accession (Uniprot) Number P48548

Gene ID 29713

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Mouse, human, pig - identical.

RRID AB_2039943.

Purity Affinity purified on immobilized antigen.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN₃.

Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4.

Isotype Rabbit IgG.

Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.

Reconstitution 25 μl, 50 μl or 0.2 ml double distilled water (DDW), depending on the sample size.

Reconstitution 0.2 ml double distilled water (DDW).

Antibody concentration after reconstitution 0.8 mg/ml.

Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).

Standard quality control of each lot Western blot analysis.

Applications: ic, if, ih, wb

May also work in: ifc*, ip*

Western blot

Rat heart membranes (1:200).
Western blot analysis of rat heart membranes:
1. Anti-KCNJ5 (K_ir3.4) Antibody (#APC-027), (1:200).
2. Anti-KCNJ5 (K_ir3.4) Antibody, preincubated with KCNJ5/Kir3.4 Blocking Peptide (#BLP-PC027).

Immunohistochemistry

Rat heart sections (1:20) (Atkinson, A.J. et al. (2013) J. Am. Heart Assoc. 2, e000246.).

Immunocytochemistry

Mouse HL-1 cells (1:500) (Nobles, M. et al. (2010) Pflugers Arch. 460, 99.).

Scientific background

K_ir3.4 is a member of the G-protein regulated inward-rectifier K⁺ (GIRK) channel subfamily which is part of an inward-rectifier K⁺ channel superfamily. The GIRK subfamily comprises four members in mammals (K_ir3.1- K_ir3.4) that present the common topology of the inward-rectifier superfamily: two transmembrane domains flanking a highly conserved pore region with the N and C-terminus located intracellularly.

K_ir3.4 and the other K_ir3 family subunits can be activated by neurotransmitters and other factors via the activation of G-protein coupled receptors. Binding of the corresponding ligand to the G-protein receptor induces the dissociation of G α-GTP from the Gβγ dimer. The latter directly binds to K_ir3 and activates the channel.

K_ir3.4 expression is largely confined to the heart, where it co-assembles with K_ir3.1 to form the prototypical muscarinic-gated K⁺ channel KACh. Indeed, knockout mice for K_ir3.4 showed impaired heart rate following vagal nerve stimulation.

A peptide toxin originating from the Apis mellifera bee venom, Tertiapin (#STT-250) was shown to be a potent blocker of K_ir3.4 containing channels (8.6 nM for the K_ir3.1/3.4 combination).⁴

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Lyophilized Powder

Image & Title:

Knockout validation of Anti-KCNJ5 (Kir3.4) Antibody in mouse heart.
Immunohistochemical staining of mice heart using Anti-KCNJ5 (K_ir3.4) Antibody (#APC-027). A. GIRK4 staining in GIRK4 KO mice atrial sections. B. GIRK4 staining in GIRK4 KO mice ventricular sections. C. GIRK4 staining in GIRK1 KO mice atrial sections and (D) ventricular sections. Staining demonstrates that the channel is localized to the outer sarcolemma and the ventricular t-tubular system. Scale bar = 10 μm.
Adapted from Liang, B. et al. (2014) with permission of the European Society of Cardiology.

For research purposes only, not for human use

Last Update: 29/10/2024

Specifications

Citations

Applications

Scientific Background

Specific Control Product

KCNJ5/Kir3.4 Blocking Peptide (#BLP-PC027) is the original antigen used for immunization during Anti-KCNJ5 (K_ir3.4) Antibody (#APC-027) generation. The blocking peptide binds and ‘blocks’ Anti-KCNJ5/Kir3.4 primary antibody, this makes it a good negative reagent control to help confirm antibody specificity in western blot and immunohistochemistry applications. This control is also often called a pre-adsorption control.

KCNJ5/Kir3.4 Blocking Peptide (#BLP-PC027)

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Certificate of Analysis

Anti-KCNJ5 (K_ir3.4) Antibody