製品の概要

  • 製品名
    Anti-MEF2A + MEF2C antibody
  • 製品の詳細
    Rabbit polyclonal to MEF2A + MEF2C
  • 特異性
    This antibody was raised against an immunogen that is predicted to recognize MEF2C, however the sequence shares high homology with MEF2A and we expect it to detect both family members.
  • アプリケーション
    適用あり: IHC-FoFr, ICC/IF, WB, Flow Cyt, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Xenopus laevis, Monkey
    交差が予測される動物種: Cow, Pig, Non human primates, Zebrafish
  • 免疫原

    Synthetic peptide within Human MEF2C aa 450 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab87425)

  • ポジティブ・コントロール

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab64644 in the following tested applications.

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

アプリケーション Abreviews 特記事項
IHC-FoFr 1/300.
ICC/IF Use a concentration of 1 - 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 51,54 kDa (predicted molecular weight: 51,54 kDa).

ab64644 detects a band at 51 kDa that corresponds to MEF2C, and a band at 54 kDa that corresponds to MEF2A.

Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

画像

  • ab64644 stained SKNSH cells. The cells were 100% MeOH fixed for 5 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64644 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150081 used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • IHC image of MEF2A/2C staining in normal mouse brain formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64644, 1µg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-MEF2A + MEF2C antibody (ab64644) at 1 µg/ml

    Lane 1 : P0 Brain (Mouse) Tissue Lysate
    Lane 2 : P7 Brain (Mouse) Tissue Lysate
    Lane 3 : P7 Brain (Rat) Tissue Lysate
    Lane 4 : P0 Brain (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 51,54 kDa
    Observed band size : MEF2A - 54,MEF2C - 51 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 60 kDa,73 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 16 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab64644 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-MEF2A + MEF2C antibody (ab64644) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Human) Tissue Lysate - fetal normal tissue
    Lane 3 : Jurkat (Human) Whole Cell Lysate (negative control)
    Lane 4 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate
    Lane 5 : Brain (Mouse) Tissue Lysate (negative control)
    Lane 6 : P0 Brain (Mouse) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 51,54 kDa
    Observed band size : MEF2A - 54,MEF2C - 51 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa,60 kDa,73 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 4 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab64644 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-MEF2A + MEF2C antibody (ab64644) at 1 µg/ml

    Lane 1 : Recombinant Human MEF2A protein (ab152519)
    Lane 2 : Human MEF2C full length protein

    Lysates/proteins at 0.1 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 51,54 kDa
    Observed band size : 55,83 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    ab64644 recognizes the full length recombinant proteins MEF2A (GST tagged) and MEF2C which have expected molecular weights of 83 and 55 kDa respectively. 

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab64644 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • IHC-P image of ab64644 stained mouse E15 tissue . The tissue was formaldehyde fixed and HIER was performed using citric acid (pH6) to permeabilise the cells. The tissue was incubated in 1%BSA at room temperature for 10 mins to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab64644, 1/100) for 2hrs at 21°C. The secondary antibody was used at a 1/200 dilution.

    See Abreview

  • IHC-FoFr image of MEF2C staining on Mouse cortex sections using ab64644 (1:300). The sections used came from animals perfused fixed with Paraformaldehyde 4% with 15% of a solution of saturated picric acid, in phosphate buffer 0.1M. Following postfixation in the same fixative overnight, the brains were cryoprotected in sucrose 30% overnight. Brains were then cut using a cryostat and the immunostainings were performed using the ‘free floating’ technique.

    See Abreview

参考文献

This product has been referenced in:
  • Ma H  et al. Direct Cardiac Reprogramming as a Novel Therapeutic Strategy for Treatment of Myocardial Infarction. Methods Mol Biol 1521:69-88 (2017). Read more (PubMed: 27910042) »
  • Di Siena S  et al. Activated c-Kit receptor in the heart promotes cardiac repair and regeneration after injury. Cell Death Dis 7:e2317 (2016). IF . Read more (PubMed: 27468693) »

See all 15 Publications for this product

レビューと Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Pig Tissue lysate - whole (Adult pig heart)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
Adult pig heart
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

投稿 Sep 29 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Zebrafish Tissue sections (heart sections)
Permeabilization
Yes - 0.1% Tween in PBS, Antigen retrieval in Citrate Buffer
Specification
heart sections
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 30°C
Fixative
Paraformaldehyde
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Abcam user community

Verified customer

投稿 Aug 28 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Zebrafish Tissue sections (Heart)
Permeabilization
Yes - PBS with 0.1% Tween-20
Specification
Heart
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jun 22 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Whole brain tissue sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
Yes - 0.05% Tween-20
Specification
Whole brain tissue sections
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

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Verified customer

投稿 May 25 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Tris-Acetate)
Loading amount
20 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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投稿 May 25 2017

Application
ChIP
Sample
Human Cell lysate - nuclear (OCI AML cells)
Negative control
Region where it is not supposed to bind
Specification
OCI AML cells
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 5 minute(s) and 0 second(s)
Specification of the cross-linking agent: PFA
Positive control
Region where it was already shown to bind (UCSC 4 regions around TSS +2600 to 5800bp)
Username

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Verified customer

投稿 Dec 12 2016

Abreviews
Application
ChIP
Sample
Rabbit Cell lysate - nuclear (heart)
Negative control
IgG
Specification
heart
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldheide
Positive control
histone H3
Username

Francesca Rusconi

Verified customer

投稿 Nov 30 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Chicken Tissue sections (Chick embryonic brain (dorsal pallium))
Specification
Chick embryonic brain (dorsal pallium)
Permeabilization
Yes - 0.1% Triton
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Dec 03 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Shyam Sundar Nandi

Verified customer

投稿 Nov 20 2014

The sequence information is related to the isoform which has been chosen to be the canonical isoform on swiss prot. Thus, I can confirm that this antibody should detect all murine as well as all human isoforms of MEF2C.

1-10 of 16 Abreviews or Q&A

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

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