HA721298

MEF2C Recombinant Rabbit Monoclonal Antibody [JE35-45]

cat.: HA721298

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Rat
Applications:	WB, IF-Cell, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	JE35-45

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 51 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human MEF2C aa 400-473.
Positive control:	THP-1 cell lysate, Rat brain tissue lysate, THP-1, human brain tissue, rat brain tissue, rat heart tissue.
Subcellular location:	Cytoplasm, Nucleus
Recommended Dilutions: WB IF-Cell IHC-P IP FC	1:2,000 1:100 1:1,000 1-2μg/sample 1:1,000
Uniprot #:	SwissProt: Q06413 Human Entrez Gene: 499497 Rat
Alternative names:	C5DELq14.3 DEL5q14.3 MADS box transcription enhancer factor 2 polypeptide C (myocyte enhancer factor 2C) MADS box transcription enhancer factor 2, polypeptide C MEF2C MEF2C_HUMAN Myocyte enhancer factor 2C Myocyte specific enhancer factor 2C Myocyte-specific enhancer factor 2C OTTHUMP00000222409 Similar to MADS box transcription enhancer factor 2 polypeptide C

Images

Fig1: Western blot analysis of MEF2C on different lysates with Rabbit anti-MEF2C antibody (HA721298) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: Jurkat cell lysate (negative)
Lane 3: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 51 kDa
Observed band size: 51/48 kDa

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 (HA721298) at 1/2,000 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.

Fig2: Immunocytochemistry analysis of THP-1 (positive) and Jurkat (negative) labeling MEF2C with Rabbit anti-MEF2C antibody (HA721298) at 1/100 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-MEF2C antibody (HA721298) at 1/100 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.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-MEF2C antibody (HA721298) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721298) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-MEF2C antibody (HA721298) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721298) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-MEF2C antibody (HA721298) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA721298) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig6: MEF2C was immunoprecipitated from 0.2 mg THP-1 cell lysate with HA721298 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA721298 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: THP-1 cell lysate (input)
Lane 2: HA721298 IP in THP-1 cell lysate
Lane 3: Rabbit IgG instead of HA721298 in THP-1 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 minutes; ECL: K1801

Fig7: Flow cytometric analysis of Jurkat (left, negative) and THP-1 (right, positive) cells labeling MEF2C.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721298, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.