bs-0061R [Primary Antibody]
Rabbit  Anti-beta-Actin (Loading Control) Polyclonal Antibody
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Host: Rabbit

Target Protein: beta-Actin (Loading Control)

IR: Immunogen Range:1-200/375

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 60

Swiss Prot: P60709

Source: Synthetic MAP peptide derived from human beta-Actin:1-200/375 

Purification: affinity purified by Protein A

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Background: 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].

Size: 50ul

Concentration: 1mg/ml

Applications: WB(1:5000-20000)
ELISA(1:5000-20000)
IHC-P(1:500-1000)
IHC-F(1:500-1000)
Flow-Cyt(1μg/Test)
IF(1:100-1000)

Cross Reactive Species: Human
Mouse
Rat
Chicken
Dog
Pig
Cow
Rabbit
Sheep
Bee
Fish
Guinea Pig
Hamster
Goat
Cat
mt,op.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample:
A549 Cell (Human) Lysate at 30 ug
Primary: Lane1: Anti-beta-Actin (bs-0061R) at 1/500 dilution
Lane2: Anti-beta-Actin (bs-0061R) at 1/1000 dilution
Lane3: Anti-beta-Actin (bs-0061R) at 1/2000 dilution
Lane4: Anti-beta-Actin (bs-0061R) at 1/4000 dilution
Lane5: Anti-beta-Actin (bs-0061R) at 1/5000 dilution
Lane6: Anti-beta-Actin (bs-0061R) at 1/8000 dilution
Lane7: Anti-beta-Actin (bs-0061R) at 1/10000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 47 kD
Sample:
A549 Cell (Human) Lysate at 30 ug
Primary: Lane1: Anti-beta-Actin (bs-0061R) at 1/500 dilution
Lane2: Anti-beta-Actin (bs-0061R) at 1/1000 dilution
Lane3: Anti-beta-Actin (bs-0061R) at 1/2000 dilution
Lane4: Anti-beta-Actin (bs-0061R) at 1/4000 dilution
Lane5: Anti-beta-Actin (bs-0061R) at 1/5000 dilution
Lane6: Anti-beta-Actin (bs-0061R) at 1/8000 dilution
Lane7: Anti-beta-Actin (bs-0061R) at 1/10000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 47 kD
Sample:
Lane1: 293T Cell Lysate at 25 ug
Lane2: A549 Cell Lysate at 25 ug
Lane3: A431 Cell Lysate at 25 ug
Primary: Anti- beta-Actin (bs-0061R) at 1/1000 and 1/5000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42kD
Observed band size: 42 kD
Sample: Lung lysate at 30ug;
Primary: Anti-beta-actin (bs-0061R) at 1:1000 dilution
Secondary: HRP conjugated Goat-Anti-Rabbit IgG(bse-0295G) at 1:3000 dilution
Predicted band size : 42kD
Observed band size : 42kD
Tissue/cell: human cervical carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Beta-actin Polyclonal Antibody, Unconjugated(bs-0061R) 1:1500, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Blank control: RSC96(blue).
Primary Antibody: Rabbit Anti-beta-Actin /FITC Conjugated antibody (bs-0061R/FITC), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG/FITC(orange) ,used under the same conditions.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice. The cells were washed twice with 1 X PBS. The cells were incubated in 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions followed by the incubated with antibody (bs-0061R/FITC, 1μg /1x10^6 cells) for 30 min on ice. Acquisition of 20,000 events was performed.
 
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