HA722023

beta Actin Recombinant Rabbit Monoclonal Antibody [PSH03-63]

cat.: HA722023

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Rabbit, Goat, Zebrafish, Dog, Monkey, Pig, Chicken, Cow, Hamster, Green monkey
Applications:	WB, IF-Cell, FC, IP
Clonality:	Monoclonal
Clone number:	PSH03-63

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 42 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within N-terminal residues of β-Actin.
Positive control:	HeLa cell lysate, HepG2 cell lysate, MCF7 cell lysate, A431 cell lysate, Jurkat cell lysate, HEK-293 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, PC-12 cell lysate, mouse testis tissue lysate, mouse spleen tissue lysate, rat testis tissue lysate, rat kidney tissue lysate, HeLa, NIH/3T3, C6.
Subcellular location:	Cytoskeleton.
Recommended Dilutions: WB IF-Cell FC IP	1:20,000-1:640,000 1:100-1:250 1:1,000 1-2μg/sample
Uniprot #:	SwissProt: P60709 Human \| P60710 Mouse \| P60711 Rat
Alternative names:	A26C1A A26C1B ACTB ACTB_HUMAN Actin beta Actin cytoplasmic 1 Actin, cytoplasmic 1, N-terminally processed Actx b actin Beta cytoskeletal actin Beta-actin BRWS1 E430023M04Rik MGC128179 PS1TP5 binding protein 1 PS1TP5BP1 β-actin β actin

Images

Fig1: Western blot analysis of beta Actin on different lysates with Rabbit anti-beta Actin antibody (HA722023) at 1/20,000 dilution.

Lane 1: HeLa cell lysate (10 µg/Lane)
Lane 2: HepG2 cell lysate (10 µg/Lane)
Lane 3: MCF7 cell lysate (10 µg/Lane)
Lane 4: A431 cell lysate (10 µg/Lane)
Lane 5: Jurkat cell lysate (10 µg/Lane)
Lane 6: HEK-293 cell lysate (10 µg/Lane)
Lane 7: RAW264.7 cell lysate (10 µg/Lane)
Lane 8: C2C12 cell lysate (10 µg/Lane)
Lane 9: PC-12 cell lysate (10 µg/Lane)
Lane 10: Mouse testis tissue lysate (10 µg/Lane)
Lane 11: Mouse spleen tissue lysate (10 µg/Lane)
Lane 12: Rat testis tissue lysate (10 µg/Lane)
Lane 13: Rat kidney tissue lysate (10 µg/Lane)

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 3 seconds;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722023) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of beta Actin on HepG2 cell lysates with Rabbit anti-beta Actin antibody (HA722023) at different dilutions.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Exposure time: 30 seconds;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722023) at different dilutions was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of HeLa cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta Actin antibody (HA722023) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig4: Flow cytometric analysis of HeLa cells labeling beta Actin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta Actin antibody (HA722023) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig6: Flow cytometric analysis of NIH/3T3 cells labeling beta Actin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig7: Immunocytochemistry analysis of L6 cells labeling beta Actin with Rabbit anti-beta Actin antibody (HA722023) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-beta Actin antibody (HA722023) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig8: Flow cytometric analysis of C6 cells labeling beta Actin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722023, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig9: beta Actin was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA722023 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722023 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 cell lysate (input)
Lane 2: HA722023 IP in NIH/3T3 cell lysate
Lane 3: Rabbit IgG instead of HA722023 in NIH/3T3 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute 2 seconds; ECL: K1801

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.