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mCherry Mouse Monoclonal Antibody [A9F2]

Usd: 350

Specification

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Catalog# HA601186

mCherry Mouse Monoclonal Antibody [A9F2]

Application

  • WB

  • IF-Cell

Reactivity

  • Species independent

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

mCherry Mouse Monoclonal Antibody [A9F2]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein within mCherry aa 1-236 / 236.

Species Reactivity

Species independent

Validated Applications

WB, IF-Cell

Conjugation

unconjugated

Clone Number

A9F2

RRID

AB_3071905

Reactivity Data

WB IF-Cell
Species Independent
Tested Tested
Tested Tested
Human
Arabidopsis Thaliana
Tested
Bovine
Tested
Cryptococcus Neoformans
Tested

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

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG2b

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:2,000

Target

Function

mCherry is a member of the mFruits family of monomeric red fluorescent proteins (mRFPs). As a RFP, mCherry was derived from DsRed of Discosoma sea anemones unlike green fluorescent proteins (GFPs) which are often derived from Aequoera victoria jellyfish.Fluorescent proteins are used to tag components in the cell, so they can be studied using fluorescence spectroscopy and fluorescence microscopy. mCherry absorbs light between 540-590 nm and emits light in the range of 550-650 nm. mCherry belongs to the group of fluorescent protein chromophores used as instruments to visualize genes and analyze their functions in experiments. Genome editing has been improved greatly through the precise insertion of these fluorescent protein tags into the genetic material of many diverse organisms. Most comparisons between the brightness and photostability of different fluorescent proteins have been made in vitro, removed from biological variables that affect protein performance in cells or organisms. It is hard to perfectly simulate cellular environments in vitro, and the difference in environment could have an effect on the brightness and photostability.

Background References

1. Sun J et al. A Glb1-2A-mCherry reporter monitors systemic aging and predicts lifespan in middle-aged mice. Nat Commun. 2022 Nov

2. Langa S et al. Evoglow-Pp1 and mCherry proteins: a dual fluorescent labeling system for lactic acid bacteria. Appl Microbiol Biotechnol. 2021 Oct

UNIPROT

SwissProt: X5DSL3 AnaplasmaMarginale

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of mCherry on different lysates with Mouse anti-mCherry antibody (<a href="/products/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/2,000 dilution.<br /><br />Lane 1: 293T-WT cell lysate (5 µg/Lane)<br />Lane 2: 293T-OE-mCherry cell lysate (5 µg/Lane)<br />Lane 3: 293T-OE-mCherry cell lysate (2.5 µg/Lane)<br />Lane 4: 293T-OE-mCherry cell lysate (1.25 µg/Lane)<br />Lane 5: 293T-OE-mCherry cell lysate (0.625 µg/Lane)<br /><br />Predicted band size: 27 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 30 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/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/150,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of mCherry on different lysates with Mouse anti-mCherry antibody (HA601186) at 1/2,000 dilution.

    Lane 1: 293T-WT cell lysate (5 µg/Lane)
    Lane 2: 293T-OE-mCherry cell lysate (5 µg/Lane)
    Lane 3: 293T-OE-mCherry cell lysate (2.5 µg/Lane)
    Lane 4: 293T-OE-mCherry cell lysate (1.25 µg/Lane)
    Lane 5: 293T-OE-mCherry cell lysate (0.625 µg/Lane)

    Predicted band size: 27 kDa
    Observed band size: 27 kDa

    Exposure time: 30 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 (HA601186) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/150,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of mCherry on mCherry recombinant protein with Mouse anti-mCherry antibody (<a href="/products/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/1,000 dilution.<br /><br />Lane 1: mCherry recombinant protein, 500ng/Lane<br />Lane 2: mCherry recombinant protein, 250ng/Lane<br />Lane 3: mCherry recombinant protein, 125ng/Lane<br />Lane 4: mCherry recombinant protein, 62.5ng/Lane<br /><br />Exposure time: 30 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/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/150,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of mCherry on mCherry recombinant protein with Mouse anti-mCherry antibody (HA601186) at 1/1,000 dilution.

    Lane 1: mCherry recombinant protein, 500ng/Lane
    Lane 2: mCherry recombinant protein, 250ng/Lane
    Lane 3: mCherry recombinant protein, 125ng/Lane
    Lane 4: mCherry recombinant protein, 62.5ng/Lane

    Exposure time: 30 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 (HA601186) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/150,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of mCherry on different lysates with Mouse anti-mCherry antibody (<a href="/products/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/5,000 dilution.<br /><br />Lane 1: 293T cell lysate<br />Lane 2: 293T transfected with mCherry-tagged cell lysate<br />Lane 3: 293T transfected with GFP-tagged cell lysate (negative)<br /><br />Lysates/proteins at 5 µg/Lane.<br /><br />Exposure time: 3 minutes; ECL: K1801;<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/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of mCherry on different lysates with Mouse anti-mCherry antibody (HA601186) at 1/5,000 dilution.

    Lane 1: 293T cell lysate
    Lane 2: 293T transfected with mCherry-tagged cell lysate
    Lane 3: 293T transfected with GFP-tagged cell lysate (negative)

    Lysates/proteins at 5 µg/Lane.

    Exposure time: 3 minutes; ECL: K1801;

    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 (HA601186) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of 293T transfected with or without mCherry cells labeling mCherry with Mouse anti-mCherry antibody (<a href="/products/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/2,000 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 Mouse anti-mCherry antibody (<a href="/products/HA601186" style="font-weight: bold;text-decoration: underline;">HA601186</a>) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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. As expected, green antibody staining is only visible in cells expressing mCherry, and green and red signals can be superimposed.

    Immunocytochemistry analysis of 293T transfected with or without mCherry cells labeling mCherry with Mouse anti-mCherry antibody (HA601186) at 1/2,000 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 Mouse anti-mCherry antibody (HA601186) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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. As expected, green antibody staining is only visible in cells expressing mCherry, and green and red signals can be superimposed.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation