Western Blot:Article Title: Plasmolipin regulates basolateral-to-apical transcytosis of ICAM-1 and leukocyte adhesion in polarized hepatic epithelial cells.
Article Snippet: .. Reagent or resource Source Identifier Mouse anti-ICAM-1 R&D Systems #BBA3; IF 1/400; IP 1/100; RRID: AB_356950 Rabbit anti-ICAM-1 Santa Cruz Biotechnology sc-7891; WB 1/1000; RRID: AB_647486 Mouse anti-ICAM-1 Santa Cruz Biotechnology sc-107; IHC 1/1000; RRID: AB_627120 Rat anti-ICAM-1 EBioscience 14–0542-81; WB 1/1000; IF 1/200; RRID: AB_529544 Rabbit anti-PLLP Our laboratory WB 1/1000; IF 1/400; IHC 1/1000; IP 1/100 Mouse anti-PLLP Our laboratory IF 1/400 Rabbit anti-ERK1/2 Santa Cruz Biotech- nology sc-94; WB 1/1000; AB_2140110 Mouse anti-tubulin Santa Cruz Biotech- nology sc-134241; WB 1/5000; RRID: AB_2009282 Rabbit anti-γ-tubulin Sigma Aldrich T3559; IF 1/2000 Mouse anti-CD3 OKT3 ATCC IF 1/50 from superna- tant generated from mouse hybridoma Mouse anti-GFP Roche 11,814,460,001; WB 1/1000; RRID: AB_390913 Rabbit anti-ZO-1 Thermo Fisher Scientific 40–2200; IF 1/500; RRID: AB_2533456 Rabbit anti-Rab11 Thermo Fisher Scientific 71–5300; WB 1/1000; IF 1/100 RRID: AB_2533987 Mouse anti-Exo70 MERCK (Millipore) MABT186 clone 70X13F3; WB 1/500 Mouse anti-IgG MERCK (SigmaAldrich) I5381; IP 1/100; RRID: AB_1163670 Rabbit anti-IgG MERCK (SigmaAldrich) I5006; IP 1/100; RRID: AB_1163659 Rabbit anti-EHD1 MERCK (SigmaAldrich) SAB2105824; WB 1/1000; RRID: AB_10740603 Rat anti-F4/80 Abcam ab6640; IF 1/1000; RRID: AB_1140040 Goat F(ab) anti mouse IgG 596 Abcam Abcam (ab6723); IF 1/100; RRID: AB_955573 Streptavidin-HRP Thermo Fisher Scientific 815–968-0747; WB 1/10000 Phalloidin-Alexa Fluor 647 Thermo Fisher Scientific A-22287; IF 1/250; RRID:AB_2620155 SiR-actin Spirochrome TimeLapse 1/5000 C. Cacho-Navas et al. 1 3 61 Page 12 of 19 Reagent or resource Source Identifier DAPI MERCK 268,298; IF 1/1000 Donkey anti-mouse Alexa Fluor 488 Thermo Fisher Scientific A-21202; IF 1/500; RRID: AB_141607 Donkey anti-mouse Alexa Fluor 555 Thermo Fisher Scientific A-31570; IF 1/500; RRID: AB_2536180 Donkey anti-mouse Alexa Fluor 647 Thermo Fisher Scientific A-31571; IF 1/500; RRID: AB_162542 Donkey anti-rabbit Alexa Fluor 488 Thermo Fisher Scientific A-21206; IF 1/500; RRID: AB_141708 Donkey anti-rabbit Alexa Fluor 555 Thermo Fisher Scientific A-31572; IF 1/500; RRID: AB_162543 Reagent or resource Source Identifier Donkey anti-rabbit Alexa Fluor 647 Thermo Fisher Scientific A-31573; IF 1/500; RRID: AB_2536183 Donkey anti-rat Alexa Fluor 488 Thermo Fisher Scientific A-21208; IF 1/500; RRID: AB_2535794 Donkey anti-mouse HRP Jackson Immunoresearch 715–035-151; WB 1/5000; RRID: AB_2340771 Donkey anti-rabbit HRP GE Healthcare NA934; WB 1/5000; RRID: AB_772206 Plasmolipin regulates basolateral‐to‐apical transcytosis of ICAM‐1 and leukocyte adhesion... 1 3 Page 13 of 19 61 .. Plasmid Backbone Origin ICAM-1-BirA* pEGFP-N1 This paper ICAM-1-GFP pEGFP-N1 Dr. F. Sánchez-Madrid (Madrid, Spain) (Barreiro et al., 2002) pSpCas9(BB)-2a-GFP PX458 Addgene; 48,138; Dr F. Zhang (Cambridge, MA) GFP pEGFP-N1 Clontech GFP-Rab11 pEGFP-C1 Dr. F. Martín-Belmonte (Madrid, Spain) (Rodriguez-Fraticelli et al., 2015) PLLP-GFP pEGFP-N1 This paper sgRNAs CRISPR/Cas9 sgRNA name 5'–3' Sequence sgRNA ICAM-1 fw CAC CGC GCA CTC CTG GTC CTG CTC G sgRNA ICAM-1 rv AAA CCG AGC AGG ACC AGG AGT GCG C sgRNA PLLP fw CAC CGG TCC GCG TGC TAA CTT TCG A sgRNA PLLP rv AAA CTC GAA AGT TAG CAC GCG GAC C
Immunohistochemistry:Article Title: Plasmolipin regulates basolateral-to-apical transcytosis of ICAM-1 and leukocyte adhesion in polarized hepatic epithelial cells.
Article Snippet: .. Reagent or resource Source Identifier Mouse anti-ICAM-1 R&D Systems #BBA3; IF 1/400; IP 1/100; RRID: AB_356950 Rabbit anti-ICAM-1 Santa Cruz Biotechnology sc-7891; WB 1/1000; RRID: AB_647486 Mouse anti-ICAM-1 Santa Cruz Biotechnology sc-107; IHC 1/1000; RRID: AB_627120 Rat anti-ICAM-1 EBioscience 14–0542-81; WB 1/1000; IF 1/200; RRID: AB_529544 Rabbit anti-PLLP Our laboratory WB 1/1000; IF 1/400; IHC 1/1000; IP 1/100 Mouse anti-PLLP Our laboratory IF 1/400 Rabbit anti-ERK1/2 Santa Cruz Biotech- nology sc-94; WB 1/1000; AB_2140110 Mouse anti-tubulin Santa Cruz Biotech- nology sc-134241; WB 1/5000; RRID: AB_2009282 Rabbit anti-γ-tubulin Sigma Aldrich T3559; IF 1/2000 Mouse anti-CD3 OKT3 ATCC IF 1/50 from superna- tant generated from mouse hybridoma Mouse anti-GFP Roche 11,814,460,001; WB 1/1000; RRID: AB_390913 Rabbit anti-ZO-1 Thermo Fisher Scientific 40–2200; IF 1/500; RRID: AB_2533456 Rabbit anti-Rab11 Thermo Fisher Scientific 71–5300; WB 1/1000; IF 1/100 RRID: AB_2533987 Mouse anti-Exo70 MERCK (Millipore) MABT186 clone 70X13F3; WB 1/500 Mouse anti-IgG MERCK (SigmaAldrich) I5381; IP 1/100; RRID: AB_1163670 Rabbit anti-IgG MERCK (SigmaAldrich) I5006; IP 1/100; RRID: AB_1163659 Rabbit anti-EHD1 MERCK (SigmaAldrich) SAB2105824; WB 1/1000; RRID: AB_10740603 Rat anti-F4/80 Abcam ab6640; IF 1/1000; RRID: AB_1140040 Goat F(ab) anti mouse IgG 596 Abcam Abcam (ab6723); IF 1/100; RRID: AB_955573 Streptavidin-HRP Thermo Fisher Scientific 815–968-0747; WB 1/10000 Phalloidin-Alexa Fluor 647 Thermo Fisher Scientific A-22287; IF 1/250; RRID:AB_2620155 SiR-actin Spirochrome TimeLapse 1/5000 C. Cacho-Navas et al. 1 3 61 Page 12 of 19 Reagent or resource Source Identifier DAPI MERCK 268,298; IF 1/1000 Donkey anti-mouse Alexa Fluor 488 Thermo Fisher Scientific A-21202; IF 1/500; RRID: AB_141607 Donkey anti-mouse Alexa Fluor 555 Thermo Fisher Scientific A-31570; IF 1/500; RRID: AB_2536180 Donkey anti-mouse Alexa Fluor 647 Thermo Fisher Scientific A-31571; IF 1/500; RRID: AB_162542 Donkey anti-rabbit Alexa Fluor 488 Thermo Fisher Scientific A-21206; IF 1/500; RRID: AB_141708 Donkey anti-rabbit Alexa Fluor 555 Thermo Fisher Scientific A-31572; IF 1/500; RRID: AB_162543 Reagent or resource Source Identifier Donkey anti-rabbit Alexa Fluor 647 Thermo Fisher Scientific A-31573; IF 1/500; RRID: AB_2536183 Donkey anti-rat Alexa Fluor 488 Thermo Fisher Scientific A-21208; IF 1/500; RRID: AB_2535794 Donkey anti-mouse HRP Jackson Immunoresearch 715–035-151; WB 1/5000; RRID: AB_2340771 Donkey anti-rabbit HRP GE Healthcare NA934; WB 1/5000; RRID: AB_772206 Plasmolipin regulates basolateral‐to‐apical transcytosis of ICAM‐1 and leukocyte adhesion... 1 3 Page 13 of 19 61 .. Plasmid Backbone Origin ICAM-1-BirA* pEGFP-N1 This paper ICAM-1-GFP pEGFP-N1 Dr. F. Sánchez-Madrid (Madrid, Spain) (Barreiro et al., 2002) pSpCas9(BB)-2a-GFP PX458 Addgene; 48,138; Dr F. Zhang (Cambridge, MA) GFP pEGFP-N1 Clontech GFP-Rab11 pEGFP-C1 Dr. F. Martín-Belmonte (Madrid, Spain) (Rodriguez-Fraticelli et al., 2015) PLLP-GFP pEGFP-N1 This paper sgRNAs CRISPR/Cas9 sgRNA name 5'–3' Sequence sgRNA ICAM-1 fw CAC CGC GCA CTC CTG GTC CTG CTC G sgRNA ICAM-1 rv AAA CCG AGC AGG ACC AGG AGT GCG C sgRNA PLLP fw CAC CGG TCC GCG TGC TAA CTT TCG A sgRNA PLLP rv AAA CTC GAA AGT TAG CAC GCG GAC C
Generated:Article Title: Plasmolipin regulates basolateral-to-apical transcytosis of ICAM-1 and leukocyte adhesion in polarized hepatic epithelial cells.
Article Snippet: .. Reagent or resource Source Identifier Mouse anti-ICAM-1 R&D Systems #BBA3; IF 1/400; IP 1/100; RRID: AB_356950 Rabbit anti-ICAM-1 Santa Cruz Biotechnology sc-7891; WB 1/1000; RRID: AB_647486 Mouse anti-ICAM-1 Santa Cruz Biotechnology sc-107; IHC 1/1000; RRID: AB_627120 Rat anti-ICAM-1 EBioscience 14–0542-81; WB 1/1000; IF 1/200; RRID: AB_529544 Rabbit anti-PLLP Our laboratory WB 1/1000; IF 1/400; IHC 1/1000; IP 1/100 Mouse anti-PLLP Our laboratory IF 1/400 Rabbit anti-ERK1/2 Santa Cruz Biotech- nology sc-94; WB 1/1000; AB_2140110 Mouse anti-tubulin Santa Cruz Biotech- nology sc-134241; WB 1/5000; RRID: AB_2009282 Rabbit anti-γ-tubulin Sigma Aldrich T3559; IF 1/2000 Mouse anti-CD3 OKT3 ATCC IF 1/50 from superna- tant generated from mouse hybridoma Mouse anti-GFP Roche 11,814,460,001; WB 1/1000; RRID: AB_390913 Rabbit anti-ZO-1 Thermo Fisher Scientific 40–2200; IF 1/500; RRID: AB_2533456 Rabbit anti-Rab11 Thermo Fisher Scientific 71–5300; WB 1/1000; IF 1/100 RRID: AB_2533987 Mouse anti-Exo70 MERCK (Millipore) MABT186 clone 70X13F3; WB 1/500 Mouse anti-IgG MERCK (SigmaAldrich) I5381; IP 1/100; RRID: AB_1163670 Rabbit anti-IgG MERCK (SigmaAldrich) I5006; IP 1/100; RRID: AB_1163659 Rabbit anti-EHD1 MERCK (SigmaAldrich) SAB2105824; WB 1/1000; RRID: AB_10740603 Rat anti-F4/80 Abcam ab6640; IF 1/1000; RRID: AB_1140040 Goat F(ab) anti mouse IgG 596 Abcam Abcam (ab6723); IF 1/100; RRID: AB_955573 Streptavidin-HRP Thermo Fisher Scientific 815–968-0747; WB 1/10000 Phalloidin-Alexa Fluor 647 Thermo Fisher Scientific A-22287; IF 1/250; RRID:AB_2620155 SiR-actin Spirochrome TimeLapse 1/5000 C. Cacho-Navas et al. 1 3 61 Page 12 of 19 Reagent or resource Source Identifier DAPI MERCK 268,298; IF 1/1000 Donkey anti-mouse Alexa Fluor 488 Thermo Fisher Scientific A-21202; IF 1/500; RRID: AB_141607 Donkey anti-mouse Alexa Fluor 555 Thermo Fisher Scientific A-31570; IF 1/500; RRID: AB_2536180 Donkey anti-mouse Alexa Fluor 647 Thermo Fisher Scientific A-31571; IF 1/500; RRID: AB_162542 Donkey anti-rabbit Alexa Fluor 488 Thermo Fisher Scientific A-21206; IF 1/500; RRID: AB_141708 Donkey anti-rabbit Alexa Fluor 555 Thermo Fisher Scientific A-31572; IF 1/500; RRID: AB_162543 Reagent or resource Source Identifier Donkey anti-rabbit Alexa Fluor 647 Thermo Fisher Scientific A-31573; IF 1/500; RRID: AB_2536183 Donkey anti-rat Alexa Fluor 488 Thermo Fisher Scientific A-21208; IF 1/500; RRID: AB_2535794 Donkey anti-mouse HRP Jackson Immunoresearch 715–035-151; WB 1/5000; RRID: AB_2340771 Donkey anti-rabbit HRP GE Healthcare NA934; WB 1/5000; RRID: AB_772206 Plasmolipin regulates basolateral‐to‐apical transcytosis of ICAM‐1 and leukocyte adhesion... 1 3 Page 13 of 19 61 .. Plasmid Backbone Origin ICAM-1-BirA* pEGFP-N1 This paper ICAM-1-GFP pEGFP-N1 Dr. F. Sánchez-Madrid (Madrid, Spain) (Barreiro et al., 2002) pSpCas9(BB)-2a-GFP PX458 Addgene; 48,138; Dr F. Zhang (Cambridge, MA) GFP pEGFP-N1 Clontech GFP-Rab11 pEGFP-C1 Dr. F. Martín-Belmonte (Madrid, Spain) (Rodriguez-Fraticelli et al., 2015) PLLP-GFP pEGFP-N1 This paper sgRNAs CRISPR/Cas9 sgRNA name 5'–3' Sequence sgRNA ICAM-1 fw CAC CGC GCA CTC CTG GTC CTG CTC G sgRNA ICAM-1 rv AAA CCG AGC AGG ACC AGG AGT GCG C sgRNA PLLP fw CAC CGG TCC GCG TGC TAA CTT TCG A sgRNA PLLP rv AAA CTC GAA AGT TAG CAC GCG GAC C
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