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mCherry / TdTomato Rabbit Polyclonal Antibody

Usd: 315

Specification

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Catalog# HA500049

mCherry / TdTomato Rabbit Polyclonal Antibody

Application

  • WB

  • ELISA

  • IF-Cell

Reactivity

  • Species independent

Conjugation

  • unconjugated

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

Overview

Product Name

mCherry / TdTomato Rabbit Polyclonal Antibody

Antibody Type

Rabbit Polyclonal Antibody

Immunogen

Recombinant protein within mCherry 1-236aa.

Product Specificity

This antibody not only recognizes mCherry, but also tdTomato.

Species Reactivity

Species independent

Validated Applications

WB, ELISA, IF-Cell

Positive Control

Recombinant mCherry protein lysates.

Conjugation

unconjugated

RRID

AB_3071149

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:10,000-1:80,000

  • ELISA

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:10,000

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. This product recognizes tdTomato.

Background References

1. Shaner, Nathan C,.et al. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature Biotechnology. 22 (12): 1567–1572.

2. Shu, Xiaokun,. et al. Novel Chromophores and Buried Charges Control Color in mFruits†,‡. Biochemistry. 45 (32): 9639–9647.

UNIPROT

SwissProt: X5DSL3 AnaplasmaMarginale

Images

  • Western blot analysis of mCherry / TdTomato on different lysates with Rabbit anti-mCherry / TdTomato antibody (<a href="/products/HA500049" style="font-weight: bold;text-decoration: underline;">HA500049</a>) at 1/10,000 dilution.<br /><br />Lane 1: 293T (Human embryonic kidney cell) transfected with mScarlet-tagged PCNA (N-terminal) cell lysate<br />Lane 2: 293T transfected with mRuby-Tomm20 (C-terminal) cell lysate<br />Lane 3: 293T transfected with mCherry-Tomm20 (C-terminal) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Exposure time: 4 seconds; 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/HA500049" style="font-weight: bold;text-decoration: underline;">HA500049</a>) at 1/10,000 dilution was used in <a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a> at room temperature. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of mCherry / TdTomato on different lysates with Rabbit anti-mCherry / TdTomato antibody (HA500049) at 1/10,000 dilution.

    Lane 1: 293T (Human embryonic kidney cell) transfected with mScarlet-tagged PCNA (N-terminal) cell lysate
    Lane 2: 293T transfected with mRuby-Tomm20 (C-terminal) cell lysate
    Lane 3: 293T transfected with mCherry-Tomm20 (C-terminal) cell lysate

    Lysates/proteins at 10 µg/Lane.

    Exposure time: 4 seconds; 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 (HA500049) at 1/10,000 dilution was used in K1803 at room temperature. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of mCherry / TdTomato on tdTomato recombinant protein with Rabbit anti-mCherry / TdTomato antibody (<a href="/products/HA500049" style="font-weight: bold;text-decoration: underline;">HA500049</a>) at 1/80,000 dilution.<br /><br />Lysates/proteins at 50 ng/Lane.<br /><br />Exposure time: 25 seconds; 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/HA500049" style="font-weight: bold;text-decoration: underline;">HA500049</a>) at 1/80,000 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 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of mCherry / TdTomato on tdTomato recombinant protein with Rabbit anti-mCherry / TdTomato antibody (HA500049) at 1/80,000 dilution.

    Lysates/proteins at 50 ng/Lane.

    Exposure time: 25 seconds; 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 (HA500049) at 1/80,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ELISA analysis of with anti-mCherry antibody. Antigen (1 μg/mL). The antigen was used as the coating antigen, and the anti-mCherry antibody was used as the capture antigen for ELISA.

    ELISA analysis of with anti-mCherry antibody. Antigen (1 μg/mL). The antigen was used as the coating antigen, and the anti-mCherry antibody was used as the capture antigen for ELISA.

  • Immunocytochemistry analysis of HeLa cells transfected with or without mCherry labeling mCherry / TdTomato with Rabbit anti-mCherry / TdTomato antibody (<a href="/products/HA500049" style="font-weight: bold;text-decoration: underline;">HA500049</a>) at 1/10,000 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-mCherry / TdTomato antibody (<a href="/products/HA500049" style="font-weight: bold;text-decoration: underline;">HA500049</a>) at 1/10,000 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.

    Immunocytochemistry analysis of HeLa cells transfected with or without mCherry labeling mCherry / TdTomato with Rabbit anti-mCherry / TdTomato antibody (HA500049) at 1/10,000 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-mCherry / TdTomato antibody (HA500049) at 1/10,000 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.

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

Citation

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