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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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Catalog #1: bsm-33128M

Applications: WB

Reactivity: Plant

Conjugate: Unconjugated

HepG2 cells 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 Transferrin(Serum Loading Control) Monoclonal Antibody(bsm-33243M)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).
25 ug total protein per lane of various lysates (see on figure) probed with Transferrin monoclonal antibody, unconjugated (bsm-33243M) at 1:5000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Catalog #1: bsm-33243m

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

Reactivity: Human

Conjugate: Unconjugated

Paraformaldehyde-fixed, paraffin embedded (rat placenta); 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 (Cyclophilin B ) Monoclonal Antibody, Unconjugated (bsm-33228M) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse placenta); 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 (Cyclophilin B ) Monoclonal Antibody, Unconjugated (bsm-33228M) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human placenta); 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 (Cyclophilin B ) Monoclonal Antibody, Unconjugated (bsm-33228M) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human small intestine); 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 (Cyclophilin B ) Monoclonal Antibody, Unconjugated (bsm-33228M) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.

Catalog #1: bsm-33228m

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

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 Beta tubulin Monoclonal Antibody, Unconjugated (bsm-33034M) at 1:400 overnight at 4\u00b0C, DAB staining.
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 Beta tubulin Monoclonal Antibody, Unconjugated (bsm-33034M) at 1:2000 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse cerebellum; 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 Beta tubulin Monoclonal Antibody, Unconjugated (bsm-33034M) at 1:2000 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse heart; 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 Beta tubulin Monoclonal Antibody, Unconjugated (bsm-33034M) at 1:200 overnight at 4\u00b0C, DAB staining.

Catalog #1: bsm-33034m

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

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

HeLa cells were stained with beta 1 Tubulin (5F7) Monoclonal Antibody, Unconjugated(bsm-33041M) at 1:50 in PBS and incubated for two hours at 37\u00b0C followed by Goat Anti-Mouse IgG (H+L) FITC conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
25 ug total protein per lane of various lysates (see on figure) probed with beta I Tubulin monoclonal antibody, unconjugated (bsm-33041M) at 1:2000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta I Tubulin Monoclonal Antibody, Unconjugated(bsm-33041M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with beta I Tubulin Monoclonal Antibody, Unconjugated(bsm-33041M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

Catalog #1: bsm-33041m

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

Reactivity: Human, Mouse

Conjugate: Unconjugated

Lane 1: Rat Pancreas tissue lysates; Lane 2: Rat Testis tissue lysates; Lane 3: Rat Thymus tissue lysates; Lane 4: Human Jurkat cell lysates; Lane 5: Human Molt-4 cell lysates; Lane 6: Human Hela cell lysates; Lane 7: Human U-2os cell lysates; Lane 8: Human THP-1 cell lysates probed with Lamin B Monoclonal Antibody, Unconjugated (bsm-33040M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Lane 1: Human Molt-4 cell lysates; Lane 2: Human Hela cell lysates; Lane 3: Human U-2os cell lysates probed with Lamin B Polyclonal Antibody, Unconjugated (bsm-33040M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.\r\n
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with lamin B1 Monoclonal Antibody, Unconjugated (ascites of bsm-33040M) at 1:300 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP 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 lamin B1 Monoclonal Antibody, Unconjugated (ascites of bsm-33040M) at 1:300 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

Catalog #1: bsm-33040m

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

Reactivity: Human

Conjugate: Unconjugated

Paraformaldehyde-fixed, paraffin embedded Rat spleen; 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 (4F8) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat 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 (4F8) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4\u00b0C, DAB staining.
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 (4F8) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat 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 (4F8) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4\u00b0C, DAB staining.

Catalog #1: bsm-33033m

Applications: WB, FCM, IHC-P

Reactivity: Human, Mouse, Rat, Hamster

Conjugate: Unconjugated

Paraformaldehyde-fixed, paraffin embedded human hepatocellular carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 20min; Antibody incubation with Serum albumin (3F4) Monoclonal Antibody, Unconjugated (bsm-0945M) at 1:500 overnight at 4\u00b0C, followed by a conjugated secondary and DAB staining.
Lane 1:Human Serum probed with Serum albumin Monoclonal Antibody, Unconjugated (bsm-0945M) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); 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; Incubation with (Human Serum albumin (Loading Control)) Monoclonal Antibody, Unconjugated (bsm-0945M) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.

Catalog #1: bsm-0945m

Applications: WB, IHC-P, IF(IHC-P)

Reactivity: Human, Mouse

Conjugate: Unconjugated

Tissue\/cell: HeLa cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal mouse serum, C-0082) at 37\u00b0C for 20 min; Incubation: Anti-Cyclin B1 Monoclonal Antibody, conjugated (bsm-33042M-AF350) 1:100, 2 hours at 37\u00b0C; PI (red, S0009) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (Histone H3\/HIST3H3 (3G1)) Monoclonal Antibody, Unconjugated (bsm-33042M ) at 1:500 overnight at 4\u00b0C, followed by a conjugated Goat Anti- Mouse IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (Histone H3\/HIST3H3 (3G1)) Monoclonal Antibody, Unconjugated (bsm-33042M ) at 1:500 overnight at 4\u00b0C, followed by a conjugated Goat Anti- Mouse IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (Histone H3\/HIST3H3 (3G1)) Monoclonal Antibody, Unconjugated (bsm-33042M ) at 1:500 overnight at 4\u00b0C, followed by a conjugated Goat Anti- Mouse IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.

Catalog #1: bsm-33042m

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

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

Jurkat cells 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 PCNA (3G1) Monoclonal Antibody(bs-R)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).
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA(Nuclear Loading Control) Monoclonal Antibody, Unconjugated(bsm-33035M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with PCNA(Nuclear Loading Control) Monoclonal Antibody, Unconjugated(bsm-33035M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Mouse, sp-0024) 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 PCNA(Nuclear Loading Control) Monoclonal Antibody, Unconjugated(bsm-33035M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Catalog #1: bsm-33035m

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

Reactivity: Human, Mouse, Rat

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