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product-image-AAA127408_FCM11.png FCM/FACS (Flow Cytometry) (Figure 3. Flow Cytometry analysis of JK cells using anti-MIS12 antibody (AAA127408).Overlay histogram showing JK cells stained with AAA127408 (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-MIS12 Antibody (AAA127408, 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 anti-Human MIS12 Polyclonal Antibody | anti-MIS12 antibody

Anti-MIS12 Antibody Picoband

Gene Names
MIS12; MTW1; hMis12; KNTC2AP; 2510025F08Rik
Reactivity
Human
Applications
ELISA, Flow Cytometry, Functional Assay, Immunofluorescence, Immunocytochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
MIS12, Antibody; Anti-MIS12 Antibody Picoband; AP-2 complex subunit beta; intraflagellar transport 88; Intraflagellar transport protein 88 homolog; Recessive polycystic kidney disease protein Tg737 homolog; Tetratricopeptide repeat protein 10; TPR repeat protein 10; IFT88; TG737; TTC10; anti-MIS12 antibody
Ordering
Host
Rabbit
Reactivity
Human
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Expressed in the heart, brain, liver, lung, kidney, skeletal muscle and pancreas.
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-MIS12 antibody
ELISA, FCM/FACS (Flow Cytometry), IF (Immunofluorescence), ICC (Immunocytochemistry), WB (Western Blot)
Immunogen
E coli-derived human MIS12 recombinant protein (Position: M1-K187).
Subcellular Localization
Centriole. Cilium basal body. Centrosome. Cilium. Cytoplasm. Flagellum.
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
Involved in primary cilium biogenesis. Also involved in autophagy since it is required for trafficking of ATG16L and the expansion of the autophagic compartment.
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/FACS (Flow Cytometry)

(Figure 3. Flow Cytometry analysis of JK cells using anti-MIS12 antibody (AAA127408).Overlay histogram showing JK cells stained with AAA127408 (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-MIS12 Antibody (AAA127408, 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.)

product-image-AAA127408_FCM11.png FCM/FACS (Flow Cytometry) (Figure 3. Flow Cytometry analysis of JK cells using anti-MIS12 antibody (AAA127408).Overlay histogram showing JK cells stained with AAA127408 (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-MIS12 Antibody (AAA127408, 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 2. IF analysis of MIS12 using anti-MIS12 antibody (AAA127408).MIS12 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5ug/mL rabbit anti-MIS12 Antibody (AAA127408) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

product-image-AAA127408_IF13.jpg IF (Immunofluorescence) (Figure 2. IF analysis of MIS12 using anti-MIS12 antibody (AAA127408).MIS12 was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5ug/mL rabbit anti-MIS12 Antibody (AAA127408) overnight at 4 degree C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.)

WB (Western Blot)

(Figure 1. Western blot analysis of MIS12 using anti-MIS12 antibody (AAA127408).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 Hela whole cell lysates,Lane 2: human 293T whole cell lysates,Lane 3: human HepG2 whole cell lysates,Lane 4: human Jurkat whole cell lysates,Lane 5: human Hacat whole cell lysates,Lane 6: human U251 whole cell lysates,Lane 7: human SH-SY5Y whole cell lysates,Lane 8: human K562 whole cell 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-MIS12 antigen affinity purified polyclonal antibody (#AAA127408) at 0.25ug/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 MIS12 at approximately 25 kDa. The expected band size for MIS12 is at 24 kDa.)

product-image-AAA127408_WB15.jpg WB (Western Blot) (Figure 1. Western blot analysis of MIS12 using anti-MIS12 antibody (AAA127408).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 Hela whole cell lysates,Lane 2: human 293T whole cell lysates,Lane 3: human HepG2 whole cell lysates,Lane 4: human Jurkat whole cell lysates,Lane 5: human Hacat whole cell lysates,Lane 6: human U251 whole cell lysates,Lane 7: human SH-SY5Y whole cell lysates,Lane 8: human K562 whole cell 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-MIS12 antigen affinity purified polyclonal antibody (#AAA127408) at 0.25ug/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 MIS12 at approximately 25 kDa. The expected band size for MIS12 is at 24 kDa.)
Related Product Information for anti-MIS12 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-MIS12 antibody
References
1. Goshima, G., Kiyomitsu, T., Yoda, K., Yanagida, M. Human centromere chromatin protein hMis12, essential for equal segregation, is independent of CENP-A loading pathway. J. Cell Biol. 160: 25-39, 2003.2. Obuse, C., Iwasaki, O., Kiyomitsu, T., Goshima, G., Toyoda, Y., Yanagida, M. A conserved Mis12 centromere complex is linked to heterochromatic HP1 and outer kinetochore protein Zwint-1. Nature Cell Biol. 6: 1135-1141, 2004.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
24,140 Da
NCBI Official Full Name
protein MIS12 homolog
NCBI Official Synonym Full Names
MIS12 kinetochore complex component
NCBI Official Symbol
MIS12
NCBI Official Synonym Symbols
MTW1; hMis12; KNTC2AP; 2510025F08Rik
NCBI Protein Information
protein MIS12 homolog; MIS12, MIND kinetochore complex component, homolog; homolog of yeast Mis12
UniProt Protein Name
Protein MIS12 homolog
UniProt Gene Name
MIS12
UniProt Entry Name
MIS12_HUMAN

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Product Notes

The MIS12 mis12 (Catalog #AAA127408) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-MIS12 Antibody Picoband reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's MIS12 can be used in a range of immunoassay formats including, but not limited to, ELISA, FCM/FACS (Flow Cytometry), IF (Immunofluorescence), ICC (Immunocytochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the MIS12 mis12 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 "MIS12, 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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