Name: Anti-Kir4.1 (KCNJ10)-ATTO Fluor-488 Antibody
Brand: Alomone Labs
SKU: APC-035-AG

Overview

Cat #: APC-035-AG

Alternative Name ATP-sensitive inward rectifier potassium channel 10, K_AB-2, BIR10, BIRK1, K_ir1.2, Potassium channel inwardly rectifying subfamily J member 10

Lyophilized Powder yes

Type: Polyclonal

Host: Rabbit

Reactivity: h, m, r

Immunogen

Peptide (C)KLEESLREQAEKEGSALSVR, corresponding to residues 356-375 of rat K_ir4.1 (Accession P49655). Intracellular, C-terminus.

Accession (Uniprot) Number P49655

Gene ID 29718

Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.

Homology Human, mouse - identical.

RRID AB_2040118.

Purity Affinity purified on immobilized antigen.

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

Isotype Rabbit IgG.

Label ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.

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

Reconstitution 50 µl double distilled water (DDW).

Antibody concentration after reconstitution 1 mg/ml.

Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).

Standard quality control of each lot Western blot analysis (unlabeled antibody, #APC-035), and immunohistochemistry (labeled antibody).

Applications: if, ih

May also work in: ic*

Immunohistochemistry

Expression of K_ir4.1 in rat cerebellum
Immunohistochemical staining of frozen sections of rat cerebellum using Anti-K_ir4.1 (KCNJ10)-ATTO Fluor-488 Antibody (#APC-035-AG) (green), (1:50). Staining is specific for Bergmann glial cells prolongations (white arrows) in the molecular layer (ML) and astrocytes (yellow arrows) in the granular layer (GL). Purkinje cell bodies are stained with fluorescent Nissl stain (red). Hoechst 33342 (blue) is used as counterstain.
Mouse brain sections (1:400) (Steiner, E. et al. (2012) Glia 60, 1646.).

References

Takumi, T. et al. (1995) J. Biol. Chem. 270, 16339.
Higashi, K. et al. (2001) Am. J. Physiol. 281, C922.
Butt, A.M. and Kalsi, A. (2006) J. Cell Mol. Med. 10, 33.

Scientific background

K_ir4.1 is a member of the inward rectifying K⁺ channel family. The family includes 15 members that are structurally and functionally different from the voltage-dependent K⁺ channels.

The family’s topology consists of two transmembrane domains that flank a single and highly conserved pore region with intracellular N- and C-termini. As is the case for the voltage-dependent K⁺ channels the functional unit for the K_ir channels is composed of four subunit that can assembly as either homo or heteromers.

K_ir channels are characterized by a K⁺ efflux that is limited by depolarizing membrane potentials thus making them essential for controlling resting membrane potential and K⁺ homeostasis.

K_ir4.1 is a member of the K_ir4 subfamily that includes one other member: K_ir4.2. K_ir4.1 can co-assemble with K_ir4.2 but also with other K_ir channels such as K_ir2.1 and K_ir5.1.

The K_ir4 subfamily has been classified as weak rectifiers with intermediate conductance.

K_ir4.1, encoded by KCNJ10, is mainly expressed in brain, specifically in glia cells, but also in retina, ear and kidney.^1,2

It has been proposed that K_ir4.1 has an essential role in glial K⁺ buffering, a process that re-uptakes the K⁺released during neuronal activity into the intracellular interstitial space. Loss of K_ir4.1 causes retinal defects and loss of endochoclear potential.³

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

Anti-K_ir4.1 (KCNJ10) Antibody (#APC-035) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot, immunoprecipitation, immunohistochemistry, and immunocytochemistry applications. It has been designed to recognize K_ir4.1 potassium channel from rat, mouse, and human samples.

Anti-K_ir4.1 (KCNJ10)-ATTO Fluor-488 Antibody (#APC-035-AG) is directly labeled with an ATTO-488 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-488 label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC. Anti-K_ir4.1 (KCNJ10)-ATTO Fluor-488 Antibody is specially suited to experiments requiring simultaneous labeling of different markers.

Certificate of Analysis SDS

For research purposes only, not for human use

Last Update: 13/02/2024

Applications

Citations

KO validation citations

Immunohistochemical staining of mouse brain sections with #APC-035. Tested in K_ir4.1^-/- mice.
Brasko, C. et al. (2017) Brain Struct. Funct. 222, 41.
Immunohistochemical staining of mouse oligodendrocyte sections with #APC-035. Tested on Kir4.1^-/- cells.
Battefeld, A. et al. (2016) Nat. Commun. 7, 11298.
Immunohistochemical staining of mouse cerebellum sections with #APC-035. Tested in K_ir4.1 conditional knockout cerebellum.
Djukic, B. et al. (2007) J. Neurosci. 27, 11354.

Immunohistochemistry citations

Mouse brain sections (1:200).
Steiner, E. et al. (2012) Glia 60, 1646.

Specifications

Scientific Background

Specific Control Products

Kir4.1/KCNJ10 Blocking Peptide (#BLP-PC035) is the original antigen used for immunization during Anti-K_ir4.1 (KCNJ10) Antibody (#APC-035) generation. The blocking peptide binds and ‘blocks’ Anti-Kir4.1/KCNJ10 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.

Kir4.1/KCNJ10 Blocking Peptide (#BLP-PC035)
Isotype controls are primary antibodies that lack specificity to the target, but match the class and type of the primary antibody used in the application. Isotype controls are used as negative controls to help differentiate non-specific background signal from specific antibody signal.
Our Rabbit IgG Isotype Control-ATTO Fluor-488 (#RIC-001-AG), was specifically developed to be used as a negative control in immunohistochemistry and surface staining flow cytometry applications, along our line of rabbit primary antibodies conjugated to the ATTO Fluor-488 fluorophore.

Rabbit IgG Isotype Control-ATTO Fluor-488 (#RIC-001-AG)

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Anti-K_ir4.1 (KCNJ10)-ATTO Fluor-488 Antibody