Direct-Blot™ HRP Primary Antibodies for Western Blotting
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World-Class Quality | Superior Customer Support | Outstanding Value Tel: 1.858.768.5800 biolegend.com 07-0108-00i Direct-Blot™ HRP Primary Antibodies for Western Blotting When performing a western blot, it is common to use a secondary antibody to detect the proteins of interest. However, when the signal doesn’t need to be amplified, there is no advantage in using a secondary conjugated antibody. Thus, we are happy to introduce our new Horse Radish Peroxidase (HRP) directly conjugated primary antibodies, Direct-Blot™, to help you perform western blots in a breeze. Our HRP directly conjugated antibodies are carefully developed and tested, so sensitivity is not compromised as compared to a traditional western blot using a conjugated secondary antibody. Advantages: • Shorter protocol • Highly sensitive • Excellent stability • Easier to multiplex • Easier to combine with Immunoprecipitation • No potential background from HRP conjugated secondary antibodies To learn more, visit: biolegend.com High sensitivity Western Blot Protocol 15 µg of total protein from either HeLa cells or Jurkat cells treated with PMA (anti- phospho ERK) was resolved by electrophoresis, transferred to nitrocellulose, and probed with different concentrations of HRP directly conjugated antibodies (Direct-Blot™) or purified monoclonal (Pur) anti-GAPDH (clone FF26A/F9 ), anti-Vimentin (clone O91D3), anti-ERK1/2 Phospho (Thr202/Tyr204) (clone 4B11B69) antibodies at a fixed concentration (0.5 µg/mL). For Direct-Blot™ antibodies, proteins were visualized using chemiluminescence detection directly. For purified antibody, proteins were visualized using a goat anti-mouse-IgG secondary antibody conjugated to HRP (Cat. No. 405306) followed by chemiluminescence detection. HeLa cells Jurkat cells
Transcript of Direct-Blot™ HRP Primary Antibodies for Western Blotting
World-Class Quality | Superior Customer Support | Outstanding Value
Tel: 1.858.768.5800biolegend.com
07-0108-00i
Direct-Blot™ HRP Primary Antibodies for Western Blotting
When performing a western blot, it is common to use a secondary antibody to detect the proteins of interest. However, when the signal doesn’t need to be amplified, there is no advantage in using a secondary conjugated antibody. Thus, we are happy to introduce our new Horse Radish Peroxidase (HRP) directly conjugated primary antibodies, Direct-Blot™, to help you perform western blots in a breeze. Our HRP directly conjugated antibodies are carefully developed and tested, so sensitivity is not compromised as compared to a traditional western blot using a conjugated secondary antibody.
Advantages:• Shorter protocol• Highly sensitive• Excellent stability• Easier to multiplex• Easier to combine with Immunoprecipitation• No potential background from HRP conjugated
secondary antibodies
To learn more, visit: biolegend.com
High sensitivity
Western Blot Protocol
15 µg of total protein from either HeLa cells or Jurkat cells treated with PMA (anti-phospho ERK) was resolved by electrophoresis, transferred to nitrocellulose, and probed with different concentrations of HRP directly conjugated antibodies (Direct-Blot™) or purified monoclonal (Pur) anti-GAPDH (clone FF26A/F9 ), anti-Vimentin (clone O91D3), anti-ERK1/2 Phospho (Thr202/Tyr204) (clone 4B11B69) antibodies at a fixed concentration (0.5 µg/mL). For Direct-Blot™ antibodies, proteins were visualized using chemiluminescence detection directly. For purified antibody, proteins were visualized using a goat anti-mouse-IgG secondary antibody conjugated to HRP (Cat. No. 405306) followed by chemiluminescence detection.
HeLa cells Jurkat cells
World-Class Quality | Superior Customer Support | Outstanding Value
Tel: 1.858.768.5800biolegend.com
07-0108-00i
Direct-Blot™ HRP Primary Antibodies For more product information, visit: biolegend.com
For the most updated list of products and to learn more, visit: biolegend.com/direct-blot
Direct-Blot™ HRP AntibodiesSpecificity Clone Reactivity Size Cat. No.β-actin 2F1-1 Human, Mouse 100 µL (20 Western Blots) 643807DYKDDDDK Tag L5 DYKDDDDK 100 µL (20 Western Blots) 637311ERK1/2 Phospho (Thr202/Tyr204) 4B11B69 Human, Mouse 100 µL (20 Western Blots) 675505GAPDH FF26A/F9 Human 100 µL (20 Western Blots) 649203HSP70 W27 Human 100 µL (20 Western Blots) 648005mCherry 8C5.5 mCherry 100 µL (20 Western Blots) 677703NF-κB p50 4D1 Human 100 µL (20 Western Blots) 616707RNA Polymerase II RPB3 1Y26 Yeast (Saccharomyces cerevisiae) 100 µL (20 Western Blots) 665005Tubulin Beta 3 (TUBB3) AA10 Mouse, Rat, Human 100 µL (20 Western Blots) 657409Vimentin O91D3 Human 100 µL (20 Western Blots) 677805
Excellent stability
10 µg of total protein was resolved by electrophoresis and transferred to a PVDF membrane. HRP directly conjugated (Direct-Blot™) antibodies were aliquoted and kept at 4° C, RT and 37° C for 2, 4, 7, and 14 days. Graphs show the percentage of activity of anti-GAPDH and anti-Vimentin at room temperature and 37° C, expressed as percentage of activity at 4° C. The band intensity quantification was done using the ImageJ software.
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