{"id":2735,"date":"2026-02-27T22:12:55","date_gmt":"2026-02-28T03:12:55","guid":{"rendered":"https:\/\/lifetein.com\/blog\/docs\/lifetein-dot-blot-protocol\/"},"modified":"2026-02-27T22:50:53","modified_gmt":"2026-02-28T03:50:53","password":"","slug":"lifetein-dot-blot-protocol","status":"publish","type":"docs","link":"https:\/\/www.lifetein.com\/blog\/docs\/lifetein-dot-blot-protocol\/","title":{"rendered":"LifeTein Dot Blot protocol"},"content":{"rendered":"

LifeTein Dot Blot protocol<\/strong><\/h1>\n

Dot Blot protocol<\/strong>
\nDot blot is similar to the Western blot technique. However, the proteins are spotted directly onto the membrane or paper for detecting and analyzing. This is a good technique to estimate protein concentration.<\/p>\n

Procedure<\/strong>
\n1. Prepare the nitrocellulose (NC) membrane.
\n2. Spot 2 \u00b5l of diluted samples onto the NC membrane and let it dry.
\n3.Blocking: Block the membrane in 1% BSA or non-fat milk in TBST (1 hr, Room Temperature).
\n4. Primary Antibody: Incubate with the primary antibody in BSA\/TBST for 1 hr at RT.
\n5. Washing: Wash three times with TBST (3 x 5 min).
\n6.Secondary antibody: Incubate with secondary antibody conjugated with HRP for 1 hr at RT.
\n7. Washing: Wash three times with TBST (15 min x 1, 5 min x 2), then once with TBS (5 min).
\n8. Developing: Incubate with ECL reagent for 1 min and expose X-ray film in the darkroom with different lengths of exposure.<\/p>\n

Reagents<\/strong>
\nTBS:
\n20 mM Tris-HCl
\n150 mM NaCl
\npH 7.5<\/p>\n

TBST:
\n0.05% Tween20 in TBS
\nBSA\/TBST
\n0.1% BSA in TBST
\nNC: Nitrocellulose membrane (BIO-RAD)<\/p>\n

<\/p>\n","protected":false},"excerpt":{"rendered":"

LifeTein Dot Blot protocol Dot Blot protocol Dot blot is similar to the Western blot technique. However, the proteins are spotted directly onto the membrane or paper for detecting and analyzing. This is a good technique to estimate protein concentration. … Continue reading →<\/span><\/a><\/p>\n","protected":false},"author":3,"featured_media":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"footnotes":""},"doc_category":[331],"doc_tag":[],"class_list":["post-2735","docs","type-docs","status-publish","hentry","doc_category-technical-protocols"],"aioseo_notices":[],"aioseo_head":"\n\t\t\n\t\n\t\n\t\n\t\n\t\n\t\n\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\n","aioseo_head_json":{"title":"LifeTein Dot Blot protocol | LifeTein Peptide Blog","description":"LifeTein Dot Blot protocol Dot Blot protocol Dot blot is similar to the Western blot technique. However, the proteins are spotted directly onto the membrane or paper for detecting and analyzing. This is a good technique to estimate protein concentration. Procedure 1. Prepare the nitrocellulose (NC) membrane. 2. Spot 2 \u00b5l of diluted samples onto","canonical_url":"https:\/\/www.lifetein.com\/blog\/docs\/lifetein-dot-blot-protocol\/","robots":"max-image-preview:large","keywords":"","webmasterTools":{"google-site-verification":"xKDW6DUiGuZ6nwglNeWlVWTgzTtMPzas13ROL63ytTc","msvalidate.01":"5B15601215D876311F84CFEFB0F72F18","miscellaneous":""},"schema":null,"og:locale":"en_US","og:site_name":"LifeTein Peptide Synthesis Blog","og:type":"article","og:title":"LifeTein Dot Blot protocol | LifeTein Peptide Blog","og:description":"LifeTein Dot Blot protocol Dot Blot protocol Dot blot is similar to the Western blot technique. However, the proteins are spotted directly onto the membrane or paper for detecting and analyzing. This is a good technique to estimate protein concentration. Procedure 1. Prepare the nitrocellulose (NC) membrane. 2. Spot 2 \u00b5l of diluted samples onto","og:url":"https:\/\/www.lifetein.com\/blog\/docs\/lifetein-dot-blot-protocol\/","fb:admins":"233983946638783","og:image":"https:\/\/www.lifetein.com\/blog\/wp-content\/uploads\/2020\/02\/LifeTein-Blog1-2-scaled.jpg","og:image:secure_url":"https:\/\/www.lifetein.com\/blog\/wp-content\/uploads\/2020\/02\/LifeTein-Blog1-2-scaled.jpg","article:published_time":"2026-02-28T03:12:55+00:00","article:modified_time":"2026-02-28T03:50:53+00:00","article:publisher":"https:\/\/www.facebook.com\/LifeTein-LLC-233983946638783\/","twitter:card":"summary_large_image","twitter:site":"@lifetein","twitter:title":"LifeTein Dot Blot protocol | LifeTein Peptide Blog","twitter:description":"LifeTein Dot Blot protocol Dot Blot protocol Dot blot is similar to the Western blot technique. However, the proteins are spotted directly onto the membrane or paper for detecting and analyzing. This is a good technique to estimate protein concentration. Procedure 1. Prepare the nitrocellulose (NC) membrane. 2. Spot 2 \u00b5l of diluted samples onto","twitter:creator":"@lifetein","twitter:image":"https:\/\/www.lifetein.com\/blog\/wp-content\/uploads\/2020\/02\/LifeTein-Blog1-2-scaled.jpg"},"aioseo_meta_data":{"post_id":"2735","title":null,"description":null,"keywords":null,"keyphrases":null,"primary_term":null,"canonical_url":null,"og_title":null,"og_description":null,"og_object_type":"default","og_image_type":"default","og_image_url":null,"og_image_width":null,"og_image_height":null,"og_image_custom_url":null,"og_image_custom_fields":null,"og_video":null,"og_custom_url":null,"og_article_section":null,"og_article_tags":null,"twitter_use_og":false,"twitter_card":"default","twitter_image_type":"default","twitter_image_url":null,"twitter_image_custom_url":null,"twitter_image_custom_fields":null,"twitter_title":null,"twitter_description":null,"schema":{"blockGraphs":[],"customGraphs":[],"default":{"data":{"Article":[],"Course":[],"Dataset":[],"FAQPage":[],"Movie":[],"Person":[],"Product":[],"ProductReview":[],"Car":[],"Recipe":[],"Service":[],"SoftwareApplication":[],"WebPage":[]},"graphName":"","isEnabled":true},"graphs":[]},"schema_type":"default","schema_type_options":null,"pillar_content":false,"robots_default":true,"robots_noindex":false,"robots_noarchive":false,"robots_nosnippet":false,"robots_nofollow":false,"robots_noimageindex":false,"robots_noodp":false,"robots_notranslate":false,"robots_max_snippet":null,"robots_max_videopreview":null,"robots_max_imagepreview":"large","priority":null,"frequency":null,"location":null,"local_seo":null,"seo_analyzer_scan_date":null,"breadcrumb_settings":null,"limit_modified_date":false,"open_ai":null,"ai":null,"created":"2026-02-28 03:12:55","updated":"2026-02-28 03:52:15"},"aioseo_breadcrumb":"

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