Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)

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Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
  • Immunoprecipitation - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
  • Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
  • Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14128(B)] to Flotillin 2/ESA
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Flotillin 2/ESA antibody [EPR14128(B)]
    See all Flotillin 2/ESA primary antibodies

  • Description

    Rabbit monoclonal [EPR14128(B)] to Flotillin 2/ESA

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293, Hela, A375 and A549 cell lysates.

  • General notes

     

     

    Previously labelled as Flotillin 2. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab181988 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB (1)
1/1000 - 1/10000. Detects a band of approximately 49 kDa (predicted molecular weight: 47 kDa).
IP
1/40.
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 49 kDa (predicted molecular weight: 47 kDa).
IP
1/40.

Target

  • Function

    May act as a scaffolding protein within caveolar membranes, functionally participating in formation of caveolae or caveolae-like vesicles. May be involved in epidermal cell adhesion and epidermal structure and function.

  • Tissue specificity

    In skin, expressed in epidermis and epidermal appendages but not in dermis. Expressed in all layers of the epidermis except the basal layer. In hair follicles, expressed in the suprabasal layer but not the basal layer. Also expressed in melanoma and carcinoma cell lines, fibroblasts and foreskin melanocytes.

  • Sequence similarities

    Belongs to the band 7/mec-2 family. Flotillin subfamily.

  • Cellular localization

    Cell membrane. Membrane > caveola. Endosome. Membrane-associated protein of caveolae.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • ECS-1 antibody
    • ECS1 antibody
    • Epidermal surface antigen 1 antibody
    • Epidermal surface antigen antibody
    • ESA antibody
    • ESA1 antibody
    • Flot2 antibody
    • FLOT2_HUMAN antibody
    • Flotillin 2 (epidermal surface antigen 1) antibody
    • Flotillin-2 antibody
    • M17S1 antibody
    • Membrane component chromosome 17 surface marker 1 antibody
    • Membrane component chromosome 17 surface marker 1 homolog antibody
    • membrane component, chromosome 17, surface marker 1 (35kD protein identified by monoclonal antibody ECS-1) antibody
    • Membrane component, chromosome 17, surface marker 1 antibody
    • REG-1 antibody
    • Reggie-1 antibody
    • Reggie-2 antibody
    see all

Images

  • Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
    Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
    All lanes : Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : FLOT2 knockout HEK293T cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 47 kDa
    Observed band size: 47 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab181988 observed at 47 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab181988 Anti-Flotillin 2/ESA antibody [EPR14128(B)] was shown to specifically react with Flotillin 2/ESA in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266847 (knockout cell lysate ab257442) was used. Wild-type and Flotillin 2/ESA knockout samples were subjected to SDS-PAGE. ab181988 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
    Immunoprecipitation - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)

    Immunoprecipitation of Hela cell lysate labeling Flotillin 2/ESA using ab181988 at a 1/40 dilution (lane 1). Lane 2 shows the negative control. Anti-Rabbit IgG (HRP) specific to the non-reduced form of IgG, was used as a secondary antibody at a dilution of 1/1500. Blocking/dilution buffer and concentration: 5% NFDM/TBST.

  • Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
    Western blot - Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
    All lanes : Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988) at 1/4000 dilution

    Lane 1 : 293 cell lysate
    Lane 2 : Hela cell lysate
    Lane 3 : A375 cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 47 kDa



    Blocking buffer and concentration: 5% NFDM/TBST

  • Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)
    Anti-Flotillin 2/ESA antibody [EPR14128(B)] (ab181988)

References (0)

Publishing research using ab181988? Please let us know so that we can cite the reference in this datasheet.

ab181988 has not yet been referenced specifically in any publications.

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Western blot abreview for Anti-Flotillin 2/ESA antibody [EPR14128(B)]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - other (Liver)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
65 µg
Specification
Liver
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Mar 18 2019

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