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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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Formalin-fixed and paraffin embedded: rat brain tissue labeled with Anti-MBP Polyclonal Antibody (bs-0380R), Unconjugated at 1:200, followed by conjugation to the secondary antibody was Goat Anti-Rabbit IgG, PE conjugated (bs-0295G-PE) at 1:200 for 40 minutes at 37\u00b0C DAPI(5ug\/ml, blue) was used to stain the cell nuclei
Formalin-fixed and paraffin embedded: rat brain tissue labeled with Anti-MBP Polyclonal Antibody (bs-0380R), Unconjugated at 1:200, followed by conjugation to the secondary antibody was Goat Anti-Rabbit IgG, PE conjugated (bs-0295G-PE) at 1:200 for 40 minutes at 37\u00b0C
Mouse brain lysates probed with MBP Polyclonal Antibody, unconjugated (bs-0380R) at 1:300 overnight at 4\u00b0C followed by a conjugated secondary antibody at 1:10000 for 60 minutes at 37\u00b0C.
A549 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 MBP Antibody(bs-0380R)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-0380R

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

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin embedded mouse intestine labeled with Rabbit Anti Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin-embedded rat brain labeled with Anti-Histone H3 Polyclonal Antibody, Unconjugated(bs-0349R) 1:200, overnight at 4\u00b0C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37\u00b0C.
Mouse splenocytes probed with Rabbit Anti-Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) (green) at 1:50 for 30 minutes followed by a FITC conjugated secondary antibody compared to unstained cells (blue), secondary only(light blue), and isotype control(orange).

Catalog #1: bs-0349R

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded rat thymus tissue labeled Anti-PCNA Polyclonal Antibody, Unconjugated (bs-0754R) at 1:200, followed by conjugation to the secondary antibody and DAB staining
293T lysates probed with PCNA Polyclonal Antibody, Unconjugated (bs-0754R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
A549 lysates probed with PCNA Polyclonal Antibody, Unconjugated (bs-0754R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
Lane 1: Human Hela cell lysates; Lane 2: Human MCF-7 cell lysates; Lane 3: Human MOLT-4 cell lysates; Lane 4: Mouse NIH\/3T3 cell lysates; Lane 5: Human HepG2 cell lysates; Lane 6: Rat Large intestine lysates probed with PCNA Polyclonal Antibody, Unconjugated (bs-0754R) 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-0754R

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

Reactivity: Human, Mouse, Rat, Chicken, Sheep, Rabbit

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: Human, Mouse, Rat

Conjugate: Unconjugated

Paraformaldehyde-fixed, paraffin embedded Mouse brain; 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 GAPDH Polyclonal Antibody, Unconjugated (bs-2188R) at 1:400 overnight at 4\u00b0C, DAB staining.
U251 lysates probed with GAPDH Polyclonal Antibody, Unconjugated (bs-2188R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
Lane 1: Hela lysates; Lane 2: MCF-7 lysates probed with GAPDH Polyclonal Antibody, Unconjugated (bs-2188R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
Mouse muscle lysates probed with GAPDH Polyclonal Antibody, Unconjugated (bs-2188R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.

Catalog #1: bs-2188R

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

Reactivity: Human, Mouse, Rat, Bovine, Pig, Chicken, Rabbit, All

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, Rabbit

Conjugate: Unconjugated

Catalog #1: bs-6640R

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

Reactivity: Human, Mouse, Rat, Rabbit

Conjugate: Unconjugated

Rat lung lysate 30ug, probed (bs-0061R) at 1:200 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, ELISA, FCM, IHC-P, IF(IHC-P), IF(IHC-F), IF(ICC)

Reactivity: Human, Mouse, Rat, Bovine, Dog, Pig, Sheep, Rabbit, Goat, Others, All

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded Human osteosarcoma labeled with Rabbit Anti-Oct-4\/POU5F1 Polyclonal Antibody, Unconjugated (bs-0830R) 1:300 followed by conjugation to the secondary antibody and DAB staining.
L1 rat brain, L2 , human colon carcinoma lysates probed (bs-0830R) at 1:200 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 90min in 37˚C. Predicted and observed band size: 41kDa.
Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Heart lysates; Lane 3: Mouse Cerebellum lysates; Lane 4: Mouse Testis lysates; Lane 5: Rat Cerebrum lysates; Lane 6: Rat Heart lysates; Lane 7: Rat Cerebellum lysates probed with Oct4 Polyclonal Antibody, Unconjugated (bs-0830R) 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-0830R

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

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated