Beta-Actin Rabbit Polyclonal Antibody (Loading Control)

• Catalog Number: orb500817
• Category: Antibodies
• Description: Beta-Actin Rabbit Polyclonal Antibody (Loading Control)
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Canine, Feline, Fish, Gallus, Guinea pig, Insect, Porcine, Rabbit, Sheep
• Reactivity: Hamster, Human, Mouse, Rat
• Isotype: IgG
• Immunogen: Synthetic MAP peptide derived from human beta-Actin (1-200/375aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:5000-50000, IHC-P=1:200-1000, IHC-F=1:200-1000, ICC/IF=1:100, IF=1:200-1000, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 42 kDa
• Target: ACTB
• UniProt ID: P60709
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Beta Actin; ACTB; Actin cytoplasmic 1; beta cytosk
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Min-Wook Hong 1,Hun Kim 1,So-Young Choi 1,Neelesh Sharma 2,*ORCID andSung-Jin Lee Effect of Gossypol on Gene Expression in Swine Granulosa Cells toxins, (2024)

PubMed:

Applications

WB

Reactivity

Porcine

Al-Yhya, Nouf et al. Pharmacological inhibition of HDAC1/3-interacting proteins induced morphological changes, and hindered the cell proliferation and migration of hepatocellular carcinoma cells Environ Sci Pollut Res Int, (2021)

PubMed: 33929667

Blank control: NIH/3T3. Primary Antibody (green line): Rabbit Anti-beta-Actin (Loading Control) antibody (orb500817), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. 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 20000 events was performed.

MCF7 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (beta-Actin) polyclonal Antibody, Unconjugated (orb500817) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Sample: Embryo Cerebrum (Mouse) Lysate at 40 ug, Du145 (Human) Lysate at 40 ug, SW480 (Human) Cell Lysate at 40 ug, U87MG (Human) Lysate at 40 ug, U251 (Human) Lysate at 40 ug, A673 (Human) Lysate at 40 ug, Lovo (Human) Lysate at 40 ug, 293FT (Human) Lysate at 40 ug, JEG-3 (Human) Lysate at 40 ug, RSC96 (Rat) Cell Lysate at 40 ug, MCF-7 (Human) Cell Lysate at 40 ug, HepG2 (Human) Lysate at 40 ug, A431 (Human) Lysate at 40 ug. Primary: Anti-beta-Actin (orb500817) at 1/2000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 42 kD.

Sample: SH-SY5Y (Human) Lysate at 40 ug, Primary: Anti-beta-Actin (orb500817) at 1/2000~1/20000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 42 kD, Observed band size: 42 kD.

Tissue/Cell: Hela cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (beta-Actin) polyclonal Antibody, Unconjugated (orb500817) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG-FITC antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.