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Product Info Summary
| SKU: | A00712-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-MCL1 Antibody Picoband®
SKU/Catalog Number
A00712-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-MCL1 Antibody Picoband® catalog # A00712-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-MCL1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00712-2)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human MCL1 recombinant protein (Position: E109-E325).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00712-2 is reactive to MCL1 in Human, Mouse, Rat
Observed Molecular Weight
40 kDa
Calculated molecular weight
37.3 kDa
Background of MCL1
MCL1, also known as myeloid cell leukemia 1, is a protein that in humans is encoded the MCL1 gene. It is mapped to 1q21.3. MCL1 is a potent multidomain antiapoptotic protein of the BCL2 family that heterodimerizes with other BCL2 family members to protect against apoptotic cell death. MCL1 as an attractive candidate for regulation of hematopoietic stem cell homeostasis that is highly expressed in hematopoietic stem cells and regulated by growth factor signals. MCL1 is a critical and specific regulator essential for ensuring the homeostasis of early hematopoietic progenitors. During mitotic arrest, MCL1 protein levels decline markedly, through a posttranslational mechanism, potentiating cell death. Phosphorylation of MCL1 directs its interaction with the tumor suppressor protein FBW7, which is the substrate-binding component of a ubiquitin ligase complex.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00712-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg /1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human Raji whole cell, human Ramos whole cell, human MCF-7 whole cell, human K562 whole cell, rat stomach tissue, rat kisney tissue
ICC/IF: PC3 cell
FCM: RT4 cell
Validation Images & Assay Conditions
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Western blot analysis of MCL1 using anti-MCL1 antibody (A00712-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Raji whole cell lysates,
Lane 2: human Ramos whole cell lysates,
Lane 3: human MCF-7 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat stomach tissue lysates,
Lane 6: rat kisney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MCL1 antigen affinity purified polyclonal antibody (Catalog # A00712-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MCL1 at approximately 40 kDa. The expected band size for MCL1 is at 28,37,40 kDa.
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IF analysis of MCL1 using anti-MCL1 antibody (A00712-2).
MCL1 was detected in an immunocytochemical section of PC3 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-MCL1 Antibody (A00712-2) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of RT4 cells using anti-MCL1 antibody (A00712-2).
Overlay histogram showing RT4 cells stained with A00712-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MCL1 Antibody (A00712-2, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Specific Publications For Anti-MCL1 Antibody Picoband® (A00712-2)
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