PRODUCT CODE: ET1606-14

Recombinant MCL1 Monoclonal Antibody (ET1606-14)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of MCL1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Raji cell lysate<br />
 Lane 2: A431 tissue lysate
  • Western blot analysis of MCL1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
 Positive control: <br />
 Lane 1: Raji cell lysate<br />
 Lane 2: A431 tissue lysate
  • Western blot analysis of MCL1 on mouse heart tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • ICC staining of MCL1 in Hela cells (green). 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 (ET1606-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of MCL1 in HepG2 cells (green). 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 (ET1606-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of MCL1 in BT-20 cells (green). 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 (ET1606-14, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-MCL1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-14, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-MCL1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-14, 1/50) for 30 minutes 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.
  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-MCL1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-14, 1/50) for 30 minutes 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.
  • Flow cytometric analysis of MCL1 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1606-14, 1/50) (blue). 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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Western blot analysis of MCL1 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1606-14, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Raji cell lysate
Lane 2: A431 tissue lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant MCL1 Monoclonal Antibody (ET1606-14)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

Raji cell lysate, a431 tissue lysate, mouse heart tissue lysates, Hela, HepG2, BT-20, human breast tissue, human kidney tissue, human pancreas tissue, Jurkat.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SI16-04

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

37/29 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

MCL1

SYNONYMS

Bcl 2 related protein EAT/mcl1 antibody; Bcl-2-like protein 3 antibody; Bcl-2-related protein EAT/mcl1 antibody; BCL2 related antibody; Bcl2-L-3 antibody; BCL2L3 antibody; EAT antibody; Induced myeloid leukemia cell differentiation protein Mcl 1 antibody; Induced myeloid leukemia cell differentiation protein Mcl-1 antibody; MCL 1 antibody; MCL1 antibody; MCL1-ES antibody; mcl1/EAT antibody; MCL1_HUMAN antibody; MCL1L antibody; MCL1S antibody; MGC104264 antibody; MGC1839 antibody; Myeloid Cell Leukemia 1 antibody; Myeloid cell leukemia ES antibody; Myeloid cell leukemia sequence 1 antibody; Myeloid cell leukemia sequence 1 BCL2 related antibody; Myeloid cell leukemia sequence 1 isoform 1 antibody; OTTHUMP00000032794 antibody; OTTHUMP00000032795 antibody; TM antibody

SEQUENCE SIMILARITIES

Belongs to the Bcl-2 family.

POST-TRANSLATIONAL MODIFICATION

Cleaved by CASP3 during apoptosis. In intact cells cleavage occurs preferentially after Asp-127, yielding a pro-apoptotic 28 kDa C-terminal fragment.; Rapidly degraded in the absence of phosphorylation on Thr-163 in the PEST region.; Phosphorylated on Ser-159, by GSK3, in response to IL3/interleukin-3 withdrawal. Phosphorylation at Ser-159 induces ubiquitination and proteasomal degradation, abrogating the anti-apoptotic activity. Treatment with taxol or okadaic acid induces phosphorylation on additional sites.; Ubiquitinated. Ubiquitination is induced by phosphorylation at Ser-159.

SUBCELLULAR LOCATION

Membrane, Cytoplasm, Mitochondrion, Nucleus.

FUNCTION

B-cell CLL/lymphoma 2 (Bcl-2) blocks cell death following a variety of stimuli and confers a death-sparing effect to certain hematopoietic cell lines following growth factor withdrawal. Myeloid cell leukemia 1 (Mcl-1) shares sequence homology with Bcl-2 and further resembles Bcl-2 in that its expression promotes cell viability. p53 and Mcl-1 demonstrate opposing effects on mitochondrial apoptosis by mediating Bcl-2 antagonist killer (Bak) activity. Mcl-1 is an important and specific regulator that is necessary for the homeostasis of early hematopoietic progenitors. Glycogen synthase kinase 3 (GSK3) controls Mcl-1 stability, which has an effect on the regulation of apoptosis by growth factors, PI 3-kinase and AKT. Mice with a deficiency of the Mcl-1 protein show a significant reduction in B and T lymphocytes similar to the effects observed in IL-7- or IL-7R-deficient mice.