Name: GIRK2/Kir3.2 Blocking Peptide
Brand: Alomone Labs
SKU: BLP-PC006

Overview

Cat #: BLP-PC006

Type: GST fusion protein

Form: Lyophilized powder

GIRK2/Kir3.2 Blocking Peptide (#BLP-PC006) is the original antigen used for immunization during Anti-GIRK2 (K_ir3.2) Antibody (#APC-006) generation. The blocking peptide binds and ‘blocks’ Anti-GIRK2/Kir3.2 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.

Certificate of Analysis SDS

Applications: wb, ihc

Application key:

WB- Western blot, IHC- Immunohistochemistry

For research purposes only. not for human use

Applications

Demonstration of Pre-adsorption control

Western blot analysis of rat brain membranes:
1. Anti-GIRK2 (K_ir3.2) Antibody (#APC-006), (1:200).
2. Anti-GIRK2 (K_ir3.2) Antibody, preincubated with GIRK2 (K_ir3.2) Blocking peptide (#BLP-PC006).
Expression of K_ir3.2 in mouse brain
Immunohistochemical staining of perfusion-fixed frozen mouse substantia nigra pars compacta sections using Anti-GIRK2 (K_ir3.2) Antibody (#APC-006), (1:400). GIRK2 staining (red) appears in cells and processes along the pars compacta of the mouse substantia nigra (horizontal arrows) and in the pars reticulata (vertical arrows). Cell nuclei are stained with DAPI (blue).
Multiplex staining of K_ir3.2 and KCNK2 in rat substantia nigra
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-GIRK2 (K_ir3.2) Antibody (#APC-006), (1:400) and Guinea pig Anti-KCNK2 (TREK-1) Antibody (#APC-047-GP), (1:120). A. K_ir3.2 staining (red) appears in cells of the substantia nigra pars compacta (SNC, arrows). B. KCNK2 (green) appears in both compacta (SNC) and reticulata (SNR) portions of the substantia nigra. C. Merge of the two images reveals co-localization in some cells (arrows), mainly in the SNC region. Cell nuclei are stained with DAPI (blue).
Mouse spinal cords (Marker, C.L. et al. (2004) J. Neurosci. 24, 2806.).
Western blot analysis of rat brain membrane (lanes 1 and 3) and mouse brain membrane (lanes 2 and 4):
1,2. Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP), (1:400).
3,4. Guinea pig Anti-GIRK2 (K_ir3.2) Antibody, preincubated with GIRK2/Kir3.2 Blocking Peptide (#BLP-PC006).

Following a screen of several secondary antibodies, the following were used for western blot analysis:
Anti-Guinea pig IgG (Sigma #A7289 or Jackson ImmunoResearch #106-035-006).
Multiplex staining of GIRK2 (K_ir3.2) and LRRK2 in mouse substantia nigra pars compacta
Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP), (1:200) and rabbit Anti-LRRK2 Antibody (#ANR-102), (1:200). A. K_ir3.2 staining (red) appears in profiles of dopaminergic neurons (arrows). B. LRRK2 staining (green) is detected in several cell bodies in the SNC. C. Merge of the two images demonstrates colocalization in several neurons (arrows). Cell nuclei are stained with DAPI (blue).
Expression of GIRK2 (K_ir3.2) in mouse brain
Immunohistochemical staining of mouse substantia nigra pars compacta (SNC) using Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP), (1:400). A. GIRK2 staining (green) appears in soma (horizontal arrows) and dendrites of dopaminergic neurons (vertical arrow). B. Nuclei staining using DAPI as the counterstain (blue). C. Merged image of panels A and B.
Expression of GIRK2 (K_ir3.2) in rat brain
Immunohistochemical staining of rat substantia nigra pars compacta (SNC) using Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP), (1:400). A. GIRK2 staining (green) appears in soma (horizontal arrows) and dendrites of dopaminergic neurons (vertical arrow). B. Nuclei staining using DAPI as the counterstain (blue). C. Merged image of panels A and B.

Properties

Sequence

GST fusion protein with the sequence ELANRAEVPLSWSVS SKLNQHAELETEEEEKNPEELTERNG, corresponding to residues 374-414 of mouse Kir3.2 (Accession P48542).

Accession (Uniprot) Number P48542

Peptide Confirmation Confirmed by DNA sequence and SDSPAGE.

Purity >95% (SDS-PAGE)

Storage Before Reconstitution Lyophilized powder can be stored intact at room temperature for two weeks. For longer periods, it should be stored at -20°C.

Reconstitution 100 μl PBS.

Concentration After Reconstitution 1.2 mg/ml.

Storage After Reconstitution -20°C.

Antigen Preadsorption Control 3 µg fusion protein per 1 µg antibody.

Standard Quality Control Of Each Lot Western blot analysis.

Specific Use it with:

Anti-GIRK2 (K_ir3.2) Antibody (#APC-006) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot, immunoprecipitation, immunocytochemistry, and immunohistochemistry applications. It has been designed to recognize K_ir3.2 from rat, mouse, and human samples.

Anti-GIRK2 (K_ir3.2) Antibody (#APC-006)
Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP, formerly AGP-013) raised in guinea pigs is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize GIRK2 from mouse, rat, and human samples. The antigen used to immunize guinea pigs is the same as Anti-GIRK2 (K_ir3.2) Antibody (#APC-006) raised in rabbit. Our line of guinea pig antibodies enables more flexibility with our products such as multiplex staining studies, immunoprecipitation, etc.

Guinea pig Anti-GIRK2 (K_ir3.2) Antibody (#APC-006-GP)