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Muscarinic Acetylcholine Receptor 2 Recombinant Rabbit Monoclonal Antibody [JE38-35]

Usd: 385

Specification

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Catalog# HA721490

Muscarinic Acetylcholine Receptor 2 Recombinant Rabbit Monoclonal Antibody [JE38-35]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

Muscarinic Acetylcholine Receptor 2 Recombinant Rabbit Monoclonal Antibody [JE38-35]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Muscarinic Acetylcholine Receptor 2 aa 140-184 / 466.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 52 kDa

Positive Control

HepG2 cell lysate, SH-SY5Y cell lysate, U-87 MG cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, human colon carcinoma tissue, mouse kidney tissue, SH-SY5Y.

Conjugation

unconjugated

Clone Number

JE38-35

RRID

AB_3072606

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IHC-P

  • 1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:500-1:1,000

Target

Function

The muscarinic acetylcholine receptor M2, also known as the cholinergic receptor, muscarinic 2, is a muscarinic acetylcholine receptor that in humans is encoded by the CHRM2 gene. Multiple alternatively spliced transcript variants have been described for this gene. M2 muscarinic receptors act via a Gi type receptor, which causes a decrease in cAMP in the cell, generally leading to inhibitory-type effects. They appear to serve as autoreceptors. In addition, they modulate G protein-coupled inwardly-rectifying potassium channels. In the heart, this contributes to a decreased heart rate. They do so by the Gβγ subunit of the G protein; Gβγ shifts the open probability of K+ channels in the membrane of the cardiac pacemaker cells, which causes an outward current of potassium, effectively hyperpolarizing the membrane, which slows down the heart rate. Few highly selective M2 agonists are available at present, although there are several non-selective muscarinic agonists that stimulate M2, and a number of selective M2 antagonists are available.

Background References

1. Boiko AS et al. Pharmacogenetics of tardive dyskinesia in schizophrenia: The role of CHRM1 and CHRM2 muscarinic receptors. World J Biol Psychiatry. 2020 Jan

Subcellular Location

Cell membrane, Postsynaptic cell membrane.

UNIPROT

SwissProt: P08172 Human

SwissProt: Q9ERZ4 Mouse

SwissProt: P10980 Rat

Synonyms

7TM receptor antibody

Acetylcholine receptor, muscarinic, 2 antibody

AChR antibody

Acm2 antibody

ACM2_HUMAN antibody

Cholinergic receptor muscarinic 2 antibody

Cholinergic receptor, muscarinic 2, cardiac antibody

Cholinergic receptor, muscarinic 2, isoform a antibody

Cholinergic receptor, muscarinic 2a antibody

CHRM 2 antibody

Expand

Images

  • Western blot analysis of Muscarinic Acetylcholine Receptor 2 on different lysates with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/1,000 dilution.<br /><br />Lane 1: HepG2 cell lysate<br />Lane 2: SH-SY5Y cell lysate<br />Lane 3: U-87 MG cell lysate<br />Lane 4: Neuro-2a cell lysate<br />Lane 5: PC-12 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 52 kDa<br /><br />Exposure time: 1 minute 17 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Muscarinic Acetylcholine Receptor 2 on different lysates with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (HA721490) at 1/1,000 dilution.

    Lane 1: HepG2 cell lysate
    Lane 2: SH-SY5Y cell lysate
    Lane 3: U-87 MG cell lysate
    Lane 4: Neuro-2a cell lysate
    Lane 5: PC-12 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 52 kDa

    Exposure time: 1 minute 17 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721490) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (HA721490) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721490) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (HA721490) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721490) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of SH-SY5Y cells labeling Muscarinic Acetylcholine Receptor 2 with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃.Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of SH-SY5Y cells labeling Muscarinic Acetylcholine Receptor 2 with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (HA721490) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Muscarinic Acetylcholine Receptor 2 antibody (HA721490) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃.Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of SH-SY5Y cells labeling Muscarinic Acetylcholine Receptor 2.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA721490" style="font-weight: bold;text-decoration: underline;">HA721490</a>, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of SH-SY5Y cells labeling Muscarinic Acetylcholine Receptor 2.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA721490, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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