Peroxidase affinipure f ab 2 fragment donkey anti rabbit igg h l

The expression of BMPER, Collagen1 (Col1), Phospho-Smad1 (p-Smad1), Smad1, Phospho-Smad3 (p-Smad3) and Smad3 were evaluated by western blotting essentially as described33 (link). For western blotting analysis, the following antibodies were used: BMPER (ab75183, 1:1000, Abcam, Cambridge, MA, USA), Collagen1 (ab21285, 1:250, Abcam), p-Smad1 (5753s, 1:1000, Cell Signaling, Danvers, MA, USA), Smad1 (9743s, 1:1000, Cell Signaling), p-Smad3 (9520p, 1:1000, Cell Signaling), Smad3 (9523s, 1:1000, Cell Signaling) and β-actin (12620, 1:1000, Cell Signaling). The secondary antibodies used were Peroxidase AffiniPure F(ab’)2 Fragment Donkey Anti-Rabbit IgG (H+L) (Code: 711-036-152, 1:5000, Jackson ImmunoResearch, West Grove, PA, USA) and Peroxidase AffiniPure Goat Anti-Mouse IgG, Fcγ Subclass 1 Specific, (Code: 115-035-205, 1:5000, Jackson ImmunoResearch,). Pierce ECL Western Blotting Substrate (Cat. # 32106, Life Technologies, Grand Island, NY, USA) was used to visualize on a ChemiDoc MP Imaging System (Bio-RAD, Philadelphia, PA, USA). Quantitative densitometric analysis relative to β-actin was used to aid clarity.

Monoclonal antibodies mouse anti-PD-1, rabbit anti-PD-L1, and mouse anti-CTLA-4 were purchased from Abcam (catalogue numbers ab52587, ab205921, and ab19792, respectively). Polyclonal rabbit anti-CD80 was purchased from MyBioSource (catalogue number MBS2522916). AffiniPure F(ab’)₂ fragment donkey anti-mouse IgG (H+L) and peroxidase AffiniPure F(ab’)₂ fragment donkey anti-rabbit IgG (H+L) were purchased from Jackson Immuno Research (catalogue numbers 715-006-150 and 711-036-152, respectively). Pierce endogenous peroxidase suppressor, TSA SuperBoost kit, and a Prolong Glass antifade mount were purchased from Thermofisher Scientific (catalogue numbers 35000, B40925 and P36980 respectively). Sodium borohydride, ATTO488 NHS ester, bovine serum albumin, and rhodamine B were purchased from Sigma Aldrich (catalogue numbers 41698-1MG-F, A2153-100G, and 234141-10G, respectively).

Anti-LXRa rabbit polyclonal(USBiological Cat # 364027), Anti-LXRb rabbit polyclonal (Abcam Cat # ab28479), Anti-FTH1 rabbit polyclonal (Cell Signaling Cat # 3998 s), Anti-SLC40A1 rabbit polyclonal (Novus Biological Cat # NBP1-21502SS), Anti-TFR1 rabbit polyclonal (Abcam Cat # ab84036), Anti-ARG2 rabbit polyclonal (ThermoFischer Cat # PA5-27987), Anti-SLC7A2 rabbit polyclonal (ThermoFischer Cat # PA5-77552), Primary- Anti F4/80 Rat Monoclonal (Abcam Cat # ab6640), Secondary- Biotinylated rabbit anti-Rat IgG (Vector Laboratories Cat # BA-4001), Mouse IgG HRP linked whole antibody (GE Healthcare Cat # NA931V), Rabbit IgG HRP linked whole antibody (GE healthcare Cat # NA934V), Anti-F4/80 antibody [CI:A3-1] (AbcamCat # ab6640), Anti-TRF1 antibody (GeneTex Cat # GTX102596), Peroxidase AffiniPure F(ab’)₂ Fragment Donkey Anti- Rabbit IgG (H + L) (Jackson ImmunoResearch Cat # 711-036-152), Anti-SLC40A1 antibody (Novus Biological Cat # NBP1-21502), Biotinylated Goat Anti-Rabbit IgG Antibody (Vector Laboratories Cat # BA-1000).