MYH14 Recombinant Rabbit Monoclonal Antibody [PSH20-41]
cat.: HA751757
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat, Monkey
Applications: WB, IHC-P, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: PSH20-41
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 228 kDa
Isotype: IgG
Immunogen: Recombinant protein within mouse MYH14 aa 1,201-1,480.
Positive control: COLO205 cell lysate, MCF7 cell lysate, HCT 116 cell lysate, HEK-293 cell lysate, COS-1 cell lysate, Mouse colon tissue lysate, Rat colon tissue lysate, MCF7, human small intestine tissue, mouse brain tissue, mouse small intestine tissue, rat brain tissue, rat small intestine tissue.
Subcellular location: Actomyosin, brush border, cytoplasm, cytosol, extracellular exosome, growth cone, membrane, myosin filament, myosin II complex, myosin II filament, stress fiber.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
  IP
1:5,000
1:1,000
1:100
1:1,000
1-2μg/sample
Uniprot #: SwissProt: Q7Z406 Human | Q6URW6 Mouse
Entrez Gene: 308572 Rat
Alternative names: 2400004E04Rik DFNA4 DKFZp667A1311 FLJ13881 FLJ43092 FP17425 II C KIAA2034 MHC16 Myh 14 MYH14 MYH14_HUMAN Myosin 14 Myosin Myosin heavy chain 14 Myosin heavy chain Myosin heavy chain non muscle IIc Myosin heavy polypeptide 14 Myosin-14 NMHC II C NMHC II-C Non muscle myosin heavy chain IIc non-muscle IIc Non-muscle myosin heavy chain IIc Nonmuscle myosin heavy chain II C OTTMUSP00000019210
Images
HA751757_1.jpg Fig1: Western blot analysis of MYH14 on different lysates with Rabbit anti-MYH14 antibody (HA751757) at 1/5,000 dilution.

Lane 1: COLO205 cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: HCT 116 cell lysate (20 µg/Lane)
Lane 4: HEK-293 cell lysate (20 µg/Lane)
Lane 5: COS-1 cell lysate (20 µg/Lane)
Lane 6: Mouse colon tissue lysate (40 µg/Lane)
Lane 7: Rat colon tissue lysate (40 µg/Lane)

Predicted band size: 228 kDa
Observed band size: 250 kDa

Exposure time: 10 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 (HA751757) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA751757_2.jpg Fig2: Immunocytochemistry analysis of MCF7 cells labeling MYH14 with Rabbit anti-MYH14 antibody (HA751757) 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-MYH14 antibody (HA751757) 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.
HA751757_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-MYH14 antibody (HA751757) 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 (HA751757) 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.
HA751757_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-MYH14 antibody (HA751757) 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 (HA751757) 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.
HA751757_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-MYH14 antibody (HA751757) 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 (HA751757) 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.
HA751757_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-MYH14 antibody (HA751757) 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 (HA751757) 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.
HA751757_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-MYH14 antibody (HA751757) 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 (HA751757) 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.
HA751757_8.jpg Fig8: Flow cytometric analysis of MCF7 cells labeling MYH14.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751757, 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).
HA751757_9.jpg Fig9: MYH14 was immunoprecipitated from 0.2 mg MCF7 cell lysate with HA751757 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751757 at 1/5,000 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: MCF7 cell lysate (input)
Lane 2: HA751757 IP in MCF7 cell lysate
Lane 3: Rabbit IgG instead of HA751757 in MCF7 cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 46 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.