VALIDATION IMAGES
Image kindly submitted by Piotr Mamczur from Wroclaw University. Immunofluorescent localization of beta actin in mouse squamous cell cancer (KLN-205 cell line) with BS-0061R (1:50) antibody and FITC-labeled secondary antibodies (1:2000). The nuclei were stained with DAPI.
Rat lung lysate 30ug, probed (bs-0061R) at 1:20000 overnight in 4˚C. Followed by conjugation to the secondary antibody (bs-0295G-HRP) at 1:3000 90min in RT. Predicted and observed band size: 42kDa.
A549 lysates (30ug) probed with different dilutions of beta Actin Polyclonal Antibody (bs-0061R), from 1:500-10,000 dilution and incubated overnight at 4˚C. Followed by IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution and 60 min incubation at 37˚C.
Lane 1: Mouse lung lysates;Lane 2: Mouse uterus lysates; Lane 3: Mouse small intestine lysates probed with beta Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Mouse Lung lysates; Lane 2: Mouse Cerebrum lysates probed with beta Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Mouse spleen cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with beta Actin Polyclonal Antibody(bs-0061R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
MCF7 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (beta-Actin) polyclonal Antibody, Unconjugated (bs-0061R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Lane 1: Mouse Thymus lysates; Lane 2: Mouse Urinary bladder lysates; Lane 3: Mouse Uterus lysates; Lane 4: Mouse Aorta lysates ; Lane 5: Mouse olfactory bulb lysates; Lane 6: Mouse Cerebellum lysates; Lane 7: Mouse Adrenal gland lysates; Lane 8: Mouse Ovary lysates; Lane 9: Mouse Ear lysates; Lane 10: U2os cell lysates; Lane 11: ASPC-1 cell lysates ; Lane 12: Mouse Vas deferens lysates; Lane 13: Mouse trachea lysates probed with beta Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:2000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Mouse Embryo Cerebrum lysates; Lane 2: Human Du145 cell lysates; Lane 3: Human SW480 cell lysates; Lane 4: Human U87MG cell lysates ; Lane 5: Human U251 cell lysates; Lane 6: Human A673 cell lysates; Lane 7: Human Lovo cell lysates; Lane 8: Human 293FT cell lysates; Lane 9: Human JEG-3 cell lysates; Lane 10: Rat RSC96 cell lysates; Lane 11: Human MCF-7 cell lysates ; Lane 12: Human HepG2 cell lysates; Lane 13: Human A431 cell lysates probed with beta Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:2000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: A549 cell lysates; Lane 2: Raw264.7 cell lysates; Lane 3: SH-SY5Y cell lysates; Lane 4: MKN45 cell lysates ; Lane 5: CHO cell lysates; Lane 6: Panc-1 cell lysates; Lane 7: 4T1 cell lysates; Lane 8: ASPC-1 cell lysates; Lane 9: H9C2 cell lysates; Lane 10: Brl-3a cell lysates; Lane 11: TT cell lysates ; Lane 12: TEV-1 cell lysates; Lane 13: EC9706 cell lysates probed with beta Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:2000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Human HL60 cell lysates; Lane 2: Human HUVEC cell lysates; Lane 3: Rat Spinal cord lysates; Lane 4: Rat Rectum lysates ; Lane 5: Rat Placenta lysates; Lane 6: Rat Lymph node lysates; Lane 7: Rat Lung lysates; Lane 8: Rat Spleen lysates; Lane 9: Human JAR cell lysates; Lane 10: Human 293T cell lysates; Lane 11: Human Jurkat cell lysates ; Lane 12: Human TM4 cell lysates probed with beta Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:2000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
NIH/3T3 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with beta Actin Polyclonal Antibody(bs-0061R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Sample:
Lymph node (Rat) Lysate at 40 ug
Primary:
Anti-beta-Actin (bs-0061R) at 1/1000~1/20000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
Sample:
Thymus (Mouse) Lysate at 40 ug
Primary:
Anti-beta-Actin (bs-0061R) at 1/1000~1/20000 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 (bs-0061R) 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
Paraformaldehyde-fixed, paraffin embedded Mouse Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta-Actin Polyclonal Antibody, Unconjugated (bs-0061R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.