MICAL1 Monoclonal antibody

MICAL1 Monoclonal Antibody for WB, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human, mouse, rat

Applications

WB, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

CloneNo.

3F5D4

Cat no : 68297-1-Ig

Synonyms

NICAL, NEDD9-interacting protein with calponin homology and LIM domains, MICAL, EC:1.14.13.225, 3F5D4

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Validation Data Gallery

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  WB analysis using 68297-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68297-1-Ig (MICAL1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with Lamin B1 Monoclonal antibody (66095-1-Ig) as loading control.

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  WB analysis of HSC-T6 using 68297-1-Ig

  HSC-T6 cells were subjected to SDS PAGE followed by western blot with 68297-1-Ig (MICAL1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of RAW 264.7 using 68297-1-Ig

  RAW 264.7 cells were subjected to SDS PAGE followed by western blot with 68297-1-Ig (MICAL1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 68297-1-Ig

  IP result of anti-MICAL1 (IP:68297-1-Ig, 4ug; Detection:68297-1-Ig 1:2000) with HeLa cells lysate 1720 ug.

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  IF Staining of HeLa using 68297-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using MICAL1 antibody (68297-1-Ig, Clone: 3F5D4 ) at dilution of 1:2000 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, red).

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  FC experiment of HeLa using 68297-1-Ig

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human MICAL1 (68297-1-Ig, Clone:3F5D4) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: A549 cells, RAW 264.7 cells, HSC-T6 cells, HeLa cells, HEK-293 cells, HepG2 cells, Jurkat cells, MOLT-4 cells, K-562 cells
• Positive IP detected in: HeLa cells
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunofluorescence (IF)/ICC: IF/ICC : 1:1000-1:4000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

68297-1-Ig targets MICAL1 in WB, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with Human, mouse, rat samples.

• Tested Reactivity: Human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: MICAL1 fusion protein Ag6578
• Full Name: microtubule associated monoxygenase, calponin and LIM domain containing 1
• Calculated Molecular Weight: 118 kDa
• Observed Molecular Weight: 120 kDa
• GenBank Accession Number: BC052983
• Gene Symbol: MICAL1
• Gene ID (NCBI): 64780
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

MICALs (Molecules Interacting with CasL) are atypical multidomain flavoenzymes with diverse cellular functions.There are three known isoforms, MICAL1, MICAL2 and MICAL3, as well as the MICAL-like proteins MICAL-L1 and MICAL-L2. MICAL1 has four conserved domains: an N-terminal flavin adenine dinucleotide (FAD) binding domain, a calponin homology (CH) domain, a Lin11, Isl-1 and Mec-3 (LIM) domain and a C-terminal coiled-coil (CC) domain. MICAL1 is reported to regulate actin stress fibers and be required for normal actin organization. It may also be involved in apoptosis through binding with NDR (nuclear Dbf2-related) kinases. This antibody specially recognizes MICAL1.