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Loading Controls

Loading Controls

 

Loading control proteins are essential for the accurate interpretation of western blot data. Loading control antibodies allow the verification of equal protein loading between samples.

Ideal loading controls are expressed constitutively and at high levels with low variability between cell lines and experimental conditions. The loading control should be at a different molecular weight than the protein of interest this allows the bands to be visually distinguishable.



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Lane 1: GFP-Tagged fusion protein (full length) Overexpression E.coli lysates(Cat#: bs-33009P); Lane 2: Control (-) E.coli lysates probed with GFP Polyclonal Antibody, Unconjugated (bs-0639R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
GFP-Tagged Fusion Protein Overexpression E.coli Lysates (Cat#: bs-33009P) probed with GFP Polyclonal Antibody, Unconjugated (bs-0639R) at 1:1000~10000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: GFP-tagged fusion protein probed with GFP Polyclonal Antibody, Unconjugated (bs-0639R) at 1:20000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Catalog #1: bs-0639R

Applications: WB, IF

Reactivity: Species independent

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded mouse colon labeled with Anti- ATP1b2\/Na+K+ATPase Polyclonal Antibody, Unconjugated (bs-1152R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
L1 rat kidney lysates L2 rat colon lysates probed with Anti ATP1b2\/Na+K+ATPase Polyclonal Antibody, Unconjugated (bs-1152R) at 1:200 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 for 90 min at RT. Predicted band 33kD. Observed band size: 33kD.
Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Human K562 cell lysates; Lane 3: Human SH-SY5Y cell lysates probed with ATPase Na+\/ K+ beta 2 Polyclonal Antibody, Unconjugated (bs-1152R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Catalog #1: bs-1152r

Applications: WB, ELISA, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity: Mouse, Rat

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded mouse embryo labeled with Anti-gamma tubulin(Centrosome Marker)Polyclonal Antibody, Unconjugated (bs-1322R) at 1:200 followed by conjugation to the secondary antibody and DAB staining.
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (gamma tubulin) polyclonal Antibody, Unconjugated (bs-1322R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Catalog #1: bs-1322R

Applications: ELISA, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity:

Conjugate: Unconjugated

Catalog #1: bs-1839R

Applications: WB, ELISA, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Rat brain lysate probed with Anti Lamin B [Nuclear Envelope Marker] Polyclonal Antibody, Unconjugated (bs-1840R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in 37˚C. Predicted band 64kD. Observed band size: 64kD
Molt-4 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Lamin B1 Polyclonal Antibody(bs-1840R)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(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Catalog #1: bs-1840R

Applications: WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity: Mouse

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded rat brain tissue labeled with Anti-BrdU(A7) Monoclonal Antibody, FITC Conjugated (bsm-0917M-FITC) at 1:200 for 40 minutes at 37\u00b0C followed by labeling Rabbit Anti-Tubulin Beta, Cy3 conjugated(bs-4511R-Cy3) 1:200, 40 minutes at 37\u00b0C
Lane 1: rat brain lysates Lane 2: rat heart lysates probed with Anti tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in 37˚C. Predicted band 50kD. Observed band size: 50kD.
Formalin-fixed and paraffin embedded rat brain labeled with Anti-tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Lane 1:Mouse heart lysates; Lane 2: Human MCF7 cell lysates probed with Rabbit Anti-Tubulin Beta Polyclonal Antibody, Unconjugated (bs-4511R) at 1:300 overnight at 4˚C. Followed by a conjugated secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at 37˚C.

Catalog #1: bs-4511R

Applications: WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

Catalog #1: bs-4963R

Applications: WB, ELISA, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Paraformaldehyde-fixed, paraffin embedded rat skeletal muscle; 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\u00b0C for 30min; Antibody incubation with COX4 (8D8) Monoclonal Antibody, Unconjugated (bsm-33037M) at 1:2000 overnight at 4\u00b0C, followed by a conjugated secondary for 20 minutes and DAB staining.
Paraformaldehyde-fixed, paraffin embedded rat skeletal muscle; 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\u00b0C for 30min; Antibody incubation with COX4 Monoclonal Antibody, Unconjugated (bsm-33037M) at 1:400 overnight at 4\u00b0C, followed by a conjugated Goat Anti-Mouse IgG antibody (bs-0296G-FITC) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded rat skeletal muscle; 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\u00b0C for 30min; Antibody incubation with COX4 Monoclonal Antibody, Unconjugated (bsm-33037M) at 1:400 overnight at 4\u00b0C, followed by a conjugated Goat Anti-Mouse IgG antibody (bs-0296G-CY3) for 90 minutes, and DAPI for nuclei staining.
Lane 1: Mouse NIH\/3T3 cell lysates; Lane 2: Mouse Cerebrum tissue lysates; Lane 3: Mouse Kidney tissue lysates; Lane 4: Mouse Heart tissue lysates; Lane 5: Rat Cerebrum tissue lysates; Lane 6: Rat Kidney tissue lysates; Lane 7: Rat Heart tissue lysates; Lane 8: Human HeLa cell lysates; Lane 9: Human Jurkat cell lysates; Lane 10: Human Raji cell lysates; Lane 11: Human 293T cell lysates; Lane 12: Human MCF-7 cell lysates probed with COX4I1(Mitochondrial Loading Control) Monoclonal Antibody, Unconjugated (bsm-33037M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Catalog #1: bsm-33037m

Applications: WB, IHC-P, IHC-F, IF

Reactivity: Human, Rat

Conjugate: Unconjugated

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.
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.
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.

Catalog #1: bs-0061r

Applications: WB, IHC-P, IHC-F, IF, Flow-Cyt, ICC/IF

Reactivity: Human, Mouse, Rat, Hamster

Conjugate: Unconjugated