Anti-MCL-1 Antibody, clone 2O7 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1430 - recombinant, expressed in HEK 293 cells
Synonym: Induced myeloid leukemia cell differentiation protein Mcl-1;Bcl-2-like protein 3;Bcl2-
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membanous/nuclear staining was observed in germinal center & non-germinal center cells, as well as in epithelium of human tonsil.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membanous/nuclear staining was observed in germinal center & non-germinal center cells, as well as in epithelium of human tonsil.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membanous/nuclear staining was observed in germinal center & non-germinal center cells, as well as in epithelium of human tonsil.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondrion, cytosol, nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondrion, cytosol, nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HCT116 cells was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondrion, cytosol, nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondrion, cytosol, nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondrion, cytosol, nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondrion, cytosol, nucleus.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Human recombinant MCL-1 protein (Left Lane) and Raji cell lysate (Right Lane) were probed with Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates MCL-1 (~35 kDa for human recombinant MCL-1 and ~40 kDa for Mcl-1 in Raji cell lysate).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 10 mg/mL of non-specific GST protein (Run 1) or human recombinant MCL-1 protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1430, Anti-MCL-1, clone 2O7 ZooMAb® Rabbit Monoclonal. The binding of this antibody to human recombinant MCL-1 protein displayed a KD value of 6.8 x 10-6, whereas its binding to non-specific GST protein displayed a KD of 1.5 x 10-3.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 2O7, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Internal |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 37.34 kDa |
| observed mol wt ~40 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_068779  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | Q07820 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Raji cell lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected MCL-1 in Raji cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected human recombinant MCL-1 protein. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected MCL-1 in human tonsil tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected MCL-1 in HCT116 and K562 cells. Affinity Binding Assay: A representative lot of this antibody bound MCL-1 Recombinant Protein with a KD of 6.8 x 10-6 in an affinity binding assay. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies.Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 17 amino acids from the internal region of human MCL-1. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 207 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects MCL-1. It targets an epitope within 17 amino acids from the internal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Induced myeloid leukemia cell differentiation protein Mcl-1 (UniProt: Q07820; also known as Bcl-2-like protein 3, Bcl2-L-3, Bcl-2-related protein EAT/mcl1, mcl1/EAT) is encoded by the MCL1 (also known as BCL2L3) gene (Gene ID: 4170) in human. MCL-1 is a Bcl-2 family protein that is involved in the regulation of apoptosis. Its expression is observed in a wide variety of cell types, in the adult and during embryonic development with tissue- and differentiation-specific variations in expression levels. It promotes cell survival by interfering at an early stage of events that lead to release of cytochrome from the mitochondria. It mediates its effects by interactions with a number of other regulators of apoptosis. It contains three BH domains (BH3, 2, and 1: aa 209-223; 304-319; and 252-272, respectively). However, it lacks BH4 domain that is present in Bcl-2 and Bcl-XL. It also contains a C-terminal transmembrane domain (aa 328-348) that serves a role to localize it to mitochondrial membrane. The N-terminal region of MCL-1 contains two PEST domains that help in its rapid turnover and short half-life. Its levels are down-regulated during apoptosis. However, it can also be cleaved by caspase 3 during apoptosis at aspartic acid 127 to yield a proapoptotic 28 kDa C-terminal fragment. MCL-1 can undergo phosphorylation at threonine 163 in the PEST region and this phosphorylation induces its ubiquitination and proteasomal degradation, thereby promoting apoptosis. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Wu, X., et al. 2020). Cell Death Dis. 11(7); Article number: 556; Michels, J., et al. (2005). Int. J. Biochem. Cell Biol. 37(2); 267-271). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |