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FCM (Flow Cytometry) (Figure 9. Flow Cytometry analysis of MCF-7 cells using anti-MLKL antibody (AAA19736).Overlay histogram showing MCF-7 cells stained with AAA19736 (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-MLKL Antibody (AAA19736, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit MLKL Polyclonal Antibody | anti-MLKL antibody

Anti-MLKL Antibody Picoband

Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Immunocytochemistry, Immunofluorescence, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
MLKL; Polyclonal Antibody; Anti-MLKL Antibody Picoband; Serine/threonine-protein kinase AtPK1/AtPK6; 70 kDa ribosomal protein S6 kinase 1 antibody; KS6B1_HUMAN antibody; p70 alpha antibody; P70 beta 1 antibody; p70 ribosomal S6 kinase alpha antibody; p70 ribosomal S6 kinase beta 1 antibody; p70 S6 kinase alpha antibody; P70 S6 Kinase antibody; p70 S6 kinase alpha 1 antibody; p70 S6 kinase alpha 2 antibody; p70 S6K antibody; p70 S6K-alpha antibody; p70 S6KA antibody; p70(S6K) alpha antibody; p70(S6K)-alpha antibody; p70-alpha antibody; p70-S6K 1 antibody; p70-S6K antibody; P70S6K antibody; P70S6K1 antibody; p70S6Kb antibody; PS6K antibody; Ribosomal protein S6 kinase 70kDa polypeptide 1 antibody; Ribosomal protein S6 kinase beta 1 antibody; Ribosomal protein S6 kinase beta-1 antibody; Ribosomal protein S6 kinase I antibody; RPS6KB1 antibody; S6K antibody; S6K-beta-1 antibody; S6K1 antibody; Serine/threonine kinase 14 alpha antibody; Serine/threonine-protein kinase 14A antibody; STK14A antibody; anti-MLKL antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Mouse, Rat
Clonality
Polyclonal
Isotype
IgG
Specificity
Expressed in all tissues.
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized. Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Applicable Applications for anti-MLKL antibody
Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml, Human, Mouse, Rat
IHC: 2-5ug/ml, Human
ICC/IF: 5ug/ml, Human
FC/FACS (Fixed): 1-3ug/1x106 cells, Human
ELISA: 0.1-0.5ug/ml
Immunogen
E coli-derived human MLKL recombinant protein (Position: R210-K464). Human MLKL shares 69.7% amino acid (aa) sequence identity with mouse MLKL.
Subcellular Localization
Cytoplasm, Nucleus
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500ug/ml.
Cross Reactivity
No cross reactivity with other proteins.
Protein Function
Downstream effector of TOR signaling pathway involved in osmotic stress response. Could be involved in the control of plant growth and development. Phosphorylates the ribosomal proteins P14, P16 and S6. Functions as a repressor of cell proliferation and required for maintenance of chromosome stability and ploidy levels through the RBR1-E2F pathway.
Preparation and Storage
At -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months.Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 9. Flow Cytometry analysis of MCF-7 cells using anti-MLKL antibody (AAA19736).Overlay histogram showing MCF-7 cells stained with AAA19736 (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-MLKL Antibody (AAA19736, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 9. Flow Cytometry analysis of MCF-7 cells using anti-MLKL antibody (AAA19736).Overlay histogram showing MCF-7 cells stained with AAA19736 (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-MLKL Antibody (AAA19736, 1ug/1x106 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IF (Immunofluorescence)

(Figure 8. IF analysis of MLKL using anti-MLKL antibody (AAA19736).MLKL was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human A549 whole cell lysates,Lane 3: human MCF-7 whole cell lysates,Lane 4: human HepG2 whole cell lysates,Lane 5: rat testis tissue lysates,Lane 6: rat liver tissue lysates,Lane 7: mouse testis tissue lysates,Lane 8: mouse liver 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-MLKL antigen affinity purified polyclonal antibody (#AAA19736) at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for MLKL at approximately 50 kDa. The expected band size for MLKL is at 54 kDa.)

IF (Immunofluorescence) (Figure 8. IF analysis of MLKL using anti-MLKL antibody (AAA19736).MLKL was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit ( epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2ug/ml rabbit anti-MLKL Antibody (AAA19736) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Lane 2: human A549 whole cell lysates,Lane 3: human MCF-7 whole cell lysates,Lane 4: human HepG2 whole cell lysates,Lane 5: rat testis tissue lysates,Lane 6: rat liver tissue lysates,Lane 7: mouse testis tissue lysates,Lane 8: mouse liver 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-MLKL antigen affinity purified polyclonal antibody (#AAA19736) at 0.5ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for MLKL at approximately 50 kDa. The expected band size for MLKL is at 54 kDa.)
Related Product Information for anti-MLKL antibody
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.
Product Categories/Family for anti-MLKL antibody
References
1. Dastani, Z., Pajukanta, P., Marcil, M., Rudzicz, N., Ruel, I., Bailey, S. D., Lee, J. C., Lemire, M., Faith, J., Platko, J., Rioux, J., Hudson, T. J., Gaudet, D., Engert, J. C., Genest, J. Fine mapping and association studies of a high-density lipoprotein cholesterol linkage region on chromosome 16 in French-Canadian subjects. Europ. J. Hum. Genet. 18: 342-347, 2010.2. Sun, L., Wang, H., Wang, Z., He, S., Chen, S., Liao, D., Wang, L., Yan, J., Liu, W., Lei, X., Wang, X. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell 148: 213-227, 2012.3. Wang, Z., Jiang, H., Chen, S., Du, F., Wang, X. The mitochondrial phosphatase PGAM5 functions at the convergence point of multiple necrotic death pathways. Cell 148: 228-243, 2012.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
54,479 Da
NCBI Official Full Name
mixed lineage kinase domain-like protein isoform 2
NCBI Official Synonym Full Names
mixed lineage kinase domain-like
NCBI Official Symbol
MLKL
NCBI Protein Information
mixed lineage kinase domain-like protein
UniProt Protein Name
Mixed lineage kinase domain-like protein
UniProt Gene Name
MLKL
UniProt Entry Name
MLKL_HUMAN

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Product Notes

The MLKL mlkl (Catalog #AAA19736) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-MLKL Antibody Picoband reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's MLKL can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS), ELISA (EIA). WB: 0.25-0.5ug/ml, Human, Mouse, Rat IHC: 2-5ug/ml, Human ICC/IF: 5ug/ml, Human FC/FACS (Fixed): 1-3ug/1x106 cells, Human ELISA: 0.1-0.5ug/ml. Researchers should empirically determine the suitability of the MLKL mlkl for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "MLKL, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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