MiTF Recombinant Rabbit Monoclonal Antibody [JF100-01]
cat.: ET1702-86
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: JF100-01
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 59 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human MiTF aa 353-520 / 526.
Positive control: SK-MEL-28 cell lysate, HeLa cell lysate, A375 cell lysate, A172 cell lysate, B16-F1 cell lysate, PC-12 cell lysate, Hela, A431, NIH/3T3, SW480.
Subcellular location: Nucleus, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  FC
1:1,000
1:50-1:100
1:50-1:100
Uniprot #: SwissProt: O75030 Human | Q08874 Mouse | O88368 Rat
Alternative names: BHLHE32 bHLHe32 Class E basic helix-loop-helix protein 32 CMM8 Homolog of mouse microphthalmia Mi Microphthalmia associated transcription factor Microphthalmia, mouse, homolog of Microphthalmia-associated transcription factor MITF MITF_HUMAN mitfa nacre WS2 WS2A z3A.1
Images
ET1702-86_1.jpg Fig1: Western blot analysis of MiTF on different lysates with Rabbit anti-MiTF antibody (ET1702-86) at 1/1,000 dilution.

Lane 1: SK-MEL-28 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)
Lane 3: A375 cell lysate (20 µg/Lane)
Lane 4: A172 cell lysate (20 µg/Lane)
Lane 5: B16-F1 cell lysate (20 µg/Lane)
Lane 6: PC-12 cell lysate (20 µg/Lane)

Predicted band size: 59 kDa
Observed band size: 50 kDa

Exposure time: 5 minutes;

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 (ET1702-86) at 1/1,000 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.
ET1702-86_2.jpg Fig2: Western blot analysis of MiTF on different lysates with Rabbit anti-MiTF antibody (ET1702-86) at 1/1,000 dilution.

Lane 1: Hela-si NT cell lysate (10 µg/Lane)
Lane 2: Hela-si MiTF cell lysate (10 µg/Lane)

Predicted band size: 59 kDa
Observed band size: 55 kDa

Exposure time: 3 minutes 26 seconds;

4-20% SDS-PAGE gel.

ET1702-86 was shown to specifically react with MiTF in Hela-si NT cells. Weakened band was observed when Hela-si MiTF sample was tested. Hela-si NT and Hela-si MiTF samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1702-86, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at 4 ℃ overnight. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1702-86_3.jpg Fig3: ICC staining of MiTF in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1702-86_4.jpg Fig4: ICC staining of MiTF in A431 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1702-86_5.jpg Fig5: ICC staining of MiTF in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1702-86_6.jpg Fig6: Flow cytometric analysis of MiTF was done on SW480 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-86, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.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”.