Anti-MEF2A + MEF2C抗体(ab64644)

概述

  • 产品名称Anti-MEF2A + MEF2C抗体
  • 描述
    兔多克隆抗体to MEF2A + MEF2C
  • 特异性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.
  • 经测试应用适用于: IHC-FoFr, ICC/IF, WB, Flow Cyt, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Xenopus laevis, Monkey
    预测可用于: Cow, Pig, Non Human Primates, Zebrafish
  • 免疫原

    Synthetic peptide corresponding to Human MEF2A/2C aa 450 to the C-terminus (C terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). immunogen correspond to MEF2C
    (Peptide available as ab87425)

  • 阳性对照

性能

应用

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.

应用 Ab评论 说明
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.

Anti-MEF2A + MEF2C antibody 图像

  • 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

Anti-MEF2A + MEF2C antibody (ab64644)参考文献

This product has been referenced in:
  • Bouttier M  et al. Alu repeats as transcriptional regulatory platforms in macrophage responses to M. tuberculosis infection. Nucleic Acids Res N/A:N/A (2016). Human . Read more (PubMed: 27604870) »
  • Oltolina F  et al. Human Cardiac Progenitor Spheroids Exhibit Enhanced Engraftment Potential. PLoS One 10:e0137999 (2015). ICC/IF ; Human . Read more (PubMed: 26375957) »

See all 12 Publications for this product

Product Wall

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

提交于 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

提交于 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

提交于 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.

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (D3 ES cells forming embryoid bodies)
Specification D3 ES cells forming embryoid bodies
Fixative Paraformaldehyde
Permeabilization Yes - trypsin-EDTA 0,1% Triton
Blocking step 0,2% BSA, 1% horse serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 0.2% · Temperature: RT°C
Username

Dr. Marta Gawor

Verified customer

提交于 Jun 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Brain)
Specification Brain
Username

Dr. Sophie Pezet

Verified customer

提交于 May 11 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Monkey Tissue sections (Spleen)
Specification Spleen
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Feb 24 2011

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (MC3T3 Osteoblasts)
Permeabilization Yes - 0.1% Triton 10 minutes
Specification MC3T3 Osteoblasts
Fixative Formaldehyde
Username

Dr. Sebastien Stephens

Verified customer

提交于 Jan 04 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Midbrain: adult)
Specification Midbrain: adult
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Aug 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (E15 embryo)
Specification E15 embryo
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Jul 27 2010

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