Home > Product > Antibody > Mouse Anti-beta-Actin (Loading Control) antibody
Beta Actin; beta-Actin; ACTB; Actin cytoplasmic 1; Actin, beta; Beta actin; beta cytoskeletal actin;A X actin like protein; ACTB; Actin cytoplasmic 1; alpha sarcomeric Actin; Actx; Beta cytoskeletal actin; Melanoma X actin; PS1TP5BP1; ACTB_HUMAN.
Cat:
SLM33036M
Species Reactivity:
Human,Mouse,Rat,(predicted: Chicken,Dog,Pig,Cow,Sheep,Fish,Guinea Pig,Hamster,Cat,)
Immunogen:
Synthetic MAP peptide derived from human beta-Actin
Format:
Liquid
Storage instructions:
Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Concentration:
1mg/ml
Clonality:
Monoclonal
Isotype:
IgG
Applications:
WB=1:5000-20000ELISA=1:5000-10000IHC-P=1:200-1000IHC-F=1:200-1000Flow-Cyt=1ug/TestICC=1:100IF=1:200-500(Paraffin sections need to do antigen repair)not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.
Host:
Mouse
Product Overview:
Sample: 293T Cell (Human) Lysate at 40 ugNIH/3T3 Cell (Mouse) Lysate at 40 ugMuscle (Mouse) Lysate at 40 ugPrimary: Anti-β-Actin (SLM33036M) at 1/5000 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 42 kDObserved band size: 42 kDSample: NIH/3T3 (Mouse) Cell Lysate at 30 ugHela (Human) Cell Lysate at 30 ugA549 (Human) Cell Lysate at 30 ugPrimary: Anti-beta-Actin (SLM33036M) at 1/1000~1/10000 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 42 kDObserved band size: 42 kDSample: Lane 1: Hela (Human) Cell Lysate at 30 ugLane 2: NIH/3T3 (Mouse) Cell Lysate at 30 ugLane 3: Lung (Rat) Lysate at 40 ugLane 4: Lung (Mouse) Lysate at 40 ugLane 5: MCF-7 (Human) Cell Lysate at 30 ugLane 6: A431 (Human) Cell Lysate at 30 ugLane 7: Stomach (Rat) Lysate at 40 ugLane 8: Stomach (Mouse) Lysate at 40 ugLane 9: Cerebrum (Rat) Lysate at 40 ugLane 10: Cerebrum (Mouse) Lysate at 40 ugPrimary: Anti-beta-Actin (SLM33036M) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 42 kDObserved band size: 42 kDSample: Lovo(Human) Cell Lysate at 30 ugPrimary: Anti- beta-Actin (SLM33036M) at 1/500 dilutionSecondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilutionPredicted band size: 42 kDObserved band size: 46 kDParaformaldehyde-fixed, paraffin embedded (mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (beta-Actin (Loading Control)) Monoclonal Antibody, Unconjugated (SLM33036M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (beta-Actin (Loading Control)) Monoclonal Antibody, Unconjugated (SLM33036M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.Blank control:Hela. Primary Antibody (green line): Mouse Anti-beta-Actin antibody (SLM33036M) Dilution: 1ug/Test; Secondary Antibody : Goat anti-Mouse IgG-FITCDilution: 0.5ug/Test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Loading Control
This gene encodes one of six different actin proteins. Actins are highly conserved proteins that are involved in cell motility, structure, and integrity. This actin is a major constituent of the contractile apparatus and one of the two nonmuscle cytoskeletal actins. [provided by RefSeq, Jul 2008].

Function:
Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.

Subunit:
Polymerization of globular actin (G-actin) leads to a structural filament (F-actin) in the form of a two-stranded helix. Each actin can bind to 4 others. Identified in a mRNP granule complex, at least composed of ACTB, ACTN4, DHX9, ERG, HNRNPA1, HNRNPA2B1, HNRNPAB, HNRNPD, HNRNPL, HNRNPR, HNRNPU, HSPA1, HSPA8, IGF2BP1, ILF2, ILF3, NCBP1, NCL, PABPC1, PABPC4, PABPN1, RPLP0, RPS3, RPS3A, RPS4X, RPS8, RPS9, SYNCRIP, TROVE2, YBX1 and untranslated mRNAs. Component of the BAF complex, which includes at least actin (ACTB), ARID1A, ARID1B/BAF250, SMARCA2, SMARCA4/BRG1, ACTL6A/BAF53, ACTL6B/BAF53B, SMARCE1/BAF57 SMARCC1/BAF155, SMARCC2/BAF170, SMARCB1/SNF5/INI1, and one or more of SMARCD1/BAF60A, SMARCD2/BAF60B, or SMARCD3/BAF60C. In muscle cells, the BAF complex also contains DPF3. Found in a complex with XPO6, Ran, ACTB and PFN1. Component of the MLL5-L complex, at least composed of MLL5, STK38, PPP1CA, PPP1CB, PPP1CC, HCFC1, ACTB and OGT. Interacts with XPO6 and EMD. Interacts with ERBB2.

Subcellular Location:
Cytoplasm. cytoskeleton.

Tissue Specificity:
Ubiquitously expressed in all eukaryotic cells.

Post-translational modifications:
ISGylated.
Oxidation of Met-44 by MICALs (MICAL1, MICAL2 or MICAL3) to form methionine sulfoxide promotes actin filament depolymerization. Methionine sulfoxide is produced stereospecifically, but it is not known whether the (S)-S-oxide or the (R)-S-oxide is produced.

DISEASE:
Defects in ACTA1 are the cause of nemaline myopathy type 3 (NEM3) [MIM:16360]. A form of nemaline myopathy. Nemaline myopathies are muscular disorders characterized by muscle weakness of varying severity and onset, and abnormal thread-or rod-like structures in muscle fibers on histologic examination. The phenotype at histological level is variable. Some patients present areas devoid of oxidative activity containg (cores) within myofibers. Core lesions are unstructured and poorly circumscribed.
Defects in ACTA1 are a cause of myopathy congenital with excess of thin myofilaments (MPCETM) [MIM:16360]. A congenital muscular disorder characterized at histological level by areas of sarcoplasm devoid of normal myofibrils and mitochondria, and replaced with dense masses of thin filaments. Central cores, rods, ragged red fibers, and necrosis are absent.

Similarity:
Belongs to the actin family.

SWISS:
P60709

Gene ID:
60

Database links:

Entrez Gene: 396526 Chicken

Entrez Gene: 60 Human

Entrez Gene: 11461 Mouse

Entrez Gene: 100009272 Rabbit

Entrez Gene: 81822 Rat

Omim: 102630 Human

SwissProt: P60706 Chicken

SwissProt: P60712 Cow

SwissProt: P60708 Horse

SwissProt: P60709 Human

SwissProt: P60710 Mouse

SwissProt: P29751 Rabbit

SwissProt: P60711 Rat

SwissProt: P60713 Sheep

Unigene: 520128 Human

Unigene: 708120 Human

Unigene: 727576 Human

Unigene: 328431 Mouse

Unigene: 391967 Mouse

Unigene: 94978 Rat



Picture

Sample:
293T Cell (Human) Lysate at 40 ug
NIH/3T3 Cell (Mouse) Lysate at 40 ug
Muscle (Mouse) Lysate at 40 ug
Primary: Anti-β-Actin (SLM33036M) at 1/5000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
Sample:
NIH/3T3 (Mouse) Cell Lysate at 30 ug
Hela (Human) Cell Lysate at 30 ug
A549 (Human) Cell Lysate at 30 ug
Primary: Anti-beta-Actin (SLM33036M) at 1/1000~1/10000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
Sample:
Lane 1: Hela (Human) Cell Lysate at 30 ug
Lane 2: NIH/3T3 (Mouse) Cell Lysate at 30 ug
Lane 3: Lung (Rat) Lysate at 40 ug
Lane 4: Lung (Mouse) Lysate at 40 ug
Lane 5: MCF-7 (Human) Cell Lysate at 30 ug
Lane 6: A431 (Human) Cell Lysate at 30 ug
Lane 7: Stomach (Rat) Lysate at 40 ug
Lane 8: Stomach (Mouse) Lysate at 40 ug
Lane 9: Cerebrum (Rat) Lysate at 40 ug
Lane 10: Cerebrum (Mouse) Lysate at 40 ug
Primary: Anti-beta-Actin (SLM33036M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
Sample:
Lovo(Human) Cell Lysate at 30 ug
Primary: Anti- beta-Actin (SLM33036M) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 46 kD
Paraformaldehyde-fixed, paraffin embedded (mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (beta-Actin (Loading Control)) Monoclonal Antibody, Unconjugated (SLM33036M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (beta-Actin (Loading Control)) Monoclonal Antibody, Unconjugated (SLM33036M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Blank control:Hela.
Primary Antibody (green line): Mouse Anti-beta-Actin antibody (SLM33036M)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-Mouse IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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