rabbit monoclonal antibody anti trim21  (Boster Bio)

Journal: Cell death & disease

Article Title: The TRIM21-FOXD1-BCL-2 axis underlies hyperglycaemic cell death and diabetic tissue damage.

doi: 10.1038/s41419-023-06355-1

Figure Lengend Snippet: Fig. 6 Hyperglycaemia induces cell apoptosis via the TRIM21-FOXD1-BCL-2 signalling axis. Chronic hyperglycaemia induces the expression of the ubiquitin E3 ligase TRIM21, which polyubiquitinates the transcription factor FOXD1 to control its protein stability, leading to hyperglycaemia-triggered downregulation of FOXD1 proteins. FOXD1 promotes the transcription of the antiapoptotic gene BCL-2 by directly binding to the BCL-2 promoter. Accordingly, hyperglycaemia-triggered degradation of FOXD1 mediated by TRIM21 shifts the balance between proapoptotic and antiapoptotic proteins and initiates apoptosis and tissue injury.

Article Snippet: The anti-HA (3724 S), anti-Flag (14793 S), anti-Myc (2276 S), anti-His (12698 S), anti-cleaved PARP (5625 S), anti-cleaved caspase 3 (9664 S), anti-cleaved caspase 8 (9496 S), anti-GAPDH (5174 S), and anti-TRIM21 (92043 S) monoclonal antibodies were purchased from Cell Signaling Technology.

Techniques: Expressing, Ubiquitin Proteomics, Control, Binding Assay

Journal: Cell reports

Article Title: TRIM21-dependent target protein ubiquitination mediates cell-free Trim-Away.

doi: 10.1016/j.celrep.2023.112125

Figure Lengend Snippet: Figure 1. Cell-free Trim-Away in Xenopus egg extracts (A) Domain architectures of full-length (FL) TRIM21 and a minimal TRIM21 construct (TRIM21R-R-PS). R, RING domain; B, B-box domain; CC, coiled-coil domain; PS, PRYSPRY domain; Fc, antibody fragment. (B) Simplified schematic of the current Trim-Away model. The target protein of interest (red) is bound by an antibody (yellow), which in turn is recognized by TRIM21 (shades of blue). Subsequent TRIM21 activation results in sequential mono- and polyubiquitination of its N terminus with K63-linked polyubiquitin, which requires the E2 ubiquitin-conjugating enzymes UBE2W and UBE2N/UBE2V2, respectively. The polyubiquitin chain on TRIM21 has been proposed to promote the proteasomal degradation of all three components. (C) Trim-Away of endogenous TFIIS in Xenopus egg extract (high-speed supernatant [HSS]) comparing recombinant human TRIM21FL and TRIM21R-R-PS (final concentrations of 1 mM). (D) Trim-Away of TFIIS in HSS using TRIM21R-R-PS in the presence of inhibitors targeting E1 enzyme (MLN7243), the proteasome (MG-262), p97 (NMS-873), or the NEDD8-activating enzyme to inhibit Cullin-RING E3 ligases (MLN4924). (E) Endogenous TFIIS was targeted by TRIM21R-R-PS in three types of Xenopus egg extract: low-speed supernatant (LSS), HSS, and nucleoplasmic extract (NPE). Note that TFIIS is more abundant in NPE than in LSS and HSS. (F) Egg extracts were supplemented with 250 nM recombinant HpaII, a bacterial DNA methyltransferase, to compare Trim-Away efficiencies using TRIM21R-R-PS

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-TRIM21 Cell Signaling Technology Cat# 92043; RRID:AB_2800177 Rabbit polyclonal anti-TFIIS Antigen: xlTFIIS full-length This paper; Pocono Rabbit Farm and Laboratory Custom Project 35585 Rabbit polyclonal anti-M.HpaII Larsen et al.34 N/A Rabbit polyclonal anti-PEX5 Romano et al.35 N/A Rabbit polyclonal anti-PNKP Stinson et al.17 N/A Rabbit polyclonal anti-FAF1 Kochenova et al.36 N/A Rabbit polyclonal anti-FAF1 Abcam Cat# ab202298 Rabbit polyclonal anti-UBXN7 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Heubes et al.37 N/A Rabbit polyclonal anti-UFD1 Heubes et al.37 N/A Rabbit polyclonal anti-AND1 Antigen: xlAND1 846-1127 This paper; Pocono Rabbit Farm and Laboratory Custom Projects 35808 and 35809 Rabbit polyclonal anti-TRAIP-C Dewar et al.38 N/A Rabbit polyclonal anti-TRAIP-I Antigen: xlTRAIP 336-351 This paper; Vivitide Custom Project 4961 Rabbit polyclonal anti-LacI Antigen: LacI-Bio This paper; BioGenes GmbH Custom Projects 28548 and 28549 Rabbit polyclonal anti-p97 Heubes et al.37 N/A Rabbit polyclonal anti-H3 Cell Signaling Technology Cat# 9715; RRID: AB_331563 Mouse monoclonal anti-Ub Santa Cruz Biotechnology Cat# sc-8017; RRID: AB_628423 Rabbit monoclonal anti-K48 polyubiquitin Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Rabbit monoclonal anti-K63 polyubiquitin Abcam Cat# ab179434; RRID:AB_2895239 Goat Anti-Rabbit IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 111-035-003; RRID: AB_2313567 Mouse Anti-Rabbit IgG (L), HRPconjugated Jackson ImmunoResearch Cat# 211-032-171; RRID: AB_2339149 Rabbit Anti-Mouse IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 315-035-003; RRID: AB_2340061 Mouse Anti-Rabbit IgG (H+L), unconjugated Jackson ImmunoResearch Cat# 211-005-109; RRID: AB_2339147 Streptavidin (HRP) Abcam Cat# ab7403 Bacterial and virus strains E. coli OverExpressTM C41(DE3) Chemically Competent Cells Sigma Cat# CMC0017 Chemicals, peptides, and recombinant proteins NEBuilder HiFi DNA Assembly Master Mix New England Biolabs Cat# E2621L IPTG Sigma Cat# I5502 DTT Bio-Rad Cat# 1610611 cOmpleteTM EDTA-free Protease Inhibitor Cocktail Roche Cat# 11873580001 (Continued on next page) Cell Reports 42, 112125, February 28, 2023 15

Techniques: Construct, Activation Assay, Ubiquitin Proteomics, Recombinant

Journal: Cell reports

Article Title: TRIM21-dependent target protein ubiquitination mediates cell-free Trim-Away.

doi: 10.1016/j.celrep.2023.112125

Figure Lengend Snippet: Figure 4. Direct polyubiquitination of target proteins is essential for their destruction (A) Schematic of experimental setup. To prevent direct target ubiquitination, proteins were chemically methylated in vitro prior to Trim-Away in egg extract. Methylation (black circle) modifies both lysine residues and the N terminus. (B and C) Trim-Away of recombinant LacI variants (final concentration of 250 nM) by TRIM21R-R-PS (B) or TRIM21FL (C) in HSS. IgG degradation fragments are highlighted. (D) Schematic of experimental setup. As an alternative means of inhibiting ubiquitination, all lysines (black squares) in target proteins were mutated to arginine (DK). Note that lysine-less mutants contain intact N termini. (E and F) Trim-Away of recombinant PEX5 variants (final concentration of 500 nM) by TRIM21R-R-PS (E) or TRIM21FL (F) in HSS. To deplete endogenous PEX5, TRIM21 and antibody were added to egg extract for 15 min (E) or 30 min (F) prior to the addition of recombinant X. laevis PEX5 to start cell-free Trim-Away at 0 min. Note that Trim-Away with TRIM21FL required an extended time course for efficient PEX5 degradation. See also Figure S3.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-TRIM21 Cell Signaling Technology Cat# 92043; RRID:AB_2800177 Rabbit polyclonal anti-TFIIS Antigen: xlTFIIS full-length This paper; Pocono Rabbit Farm and Laboratory Custom Project 35585 Rabbit polyclonal anti-M.HpaII Larsen et al.34 N/A Rabbit polyclonal anti-PEX5 Romano et al.35 N/A Rabbit polyclonal anti-PNKP Stinson et al.17 N/A Rabbit polyclonal anti-FAF1 Kochenova et al.36 N/A Rabbit polyclonal anti-FAF1 Abcam Cat# ab202298 Rabbit polyclonal anti-UBXN7 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Heubes et al.37 N/A Rabbit polyclonal anti-UFD1 Heubes et al.37 N/A Rabbit polyclonal anti-AND1 Antigen: xlAND1 846-1127 This paper; Pocono Rabbit Farm and Laboratory Custom Projects 35808 and 35809 Rabbit polyclonal anti-TRAIP-C Dewar et al.38 N/A Rabbit polyclonal anti-TRAIP-I Antigen: xlTRAIP 336-351 This paper; Vivitide Custom Project 4961 Rabbit polyclonal anti-LacI Antigen: LacI-Bio This paper; BioGenes GmbH Custom Projects 28548 and 28549 Rabbit polyclonal anti-p97 Heubes et al.37 N/A Rabbit polyclonal anti-H3 Cell Signaling Technology Cat# 9715; RRID: AB_331563 Mouse monoclonal anti-Ub Santa Cruz Biotechnology Cat# sc-8017; RRID: AB_628423 Rabbit monoclonal anti-K48 polyubiquitin Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Rabbit monoclonal anti-K63 polyubiquitin Abcam Cat# ab179434; RRID:AB_2895239 Goat Anti-Rabbit IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 111-035-003; RRID: AB_2313567 Mouse Anti-Rabbit IgG (L), HRPconjugated Jackson ImmunoResearch Cat# 211-032-171; RRID: AB_2339149 Rabbit Anti-Mouse IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 315-035-003; RRID: AB_2340061 Mouse Anti-Rabbit IgG (H+L), unconjugated Jackson ImmunoResearch Cat# 211-005-109; RRID: AB_2339147 Streptavidin (HRP) Abcam Cat# ab7403 Bacterial and virus strains E. coli OverExpressTM C41(DE3) Chemically Competent Cells Sigma Cat# CMC0017 Chemicals, peptides, and recombinant proteins NEBuilder HiFi DNA Assembly Master Mix New England Biolabs Cat# E2621L IPTG Sigma Cat# I5502 DTT Bio-Rad Cat# 1610611 cOmpleteTM EDTA-free Protease Inhibitor Cocktail Roche Cat# 11873580001 (Continued on next page) Cell Reports 42, 112125, February 28, 2023 15

Techniques: Ubiquitin Proteomics, Methylation, In Vitro, Recombinant, Concentration Assay

Journal: Cell reports

Article Title: TRIM21-dependent target protein ubiquitination mediates cell-free Trim-Away.

doi: 10.1016/j.celrep.2023.112125

Figure Lengend Snippet: Figure 5. Direct target polyubiquitination is sufficient for Trim-Away (A) Schematic of experimental setup. To prevent direct ubiquitination of TRIM21 or antibody, reductive methylation (black circle) was performed in vitro prior to Trim-Away in egg extract. Note that this procedure inhibits ubiquitination of both the N terminus and lysine residues. (B and C) Cell-free Trim-Away of recombinant unmethylated HpaII (final concentration of 250 nM) by HpaII antibody and TRIM21R-R-PS (B) or TRIM21FL (C). The methylation state of HpaII antibody and TRIM21 variant is indicated. Trim-Away with TRIM21FL required an extended time course for efficient HpaII degradation. Note that methylation of TRIM21FL resulted in an even stronger non-specific band (asterisks), which is already present at the beginning of the Trim-Away reaction. See also Figure S4.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-TRIM21 Cell Signaling Technology Cat# 92043; RRID:AB_2800177 Rabbit polyclonal anti-TFIIS Antigen: xlTFIIS full-length This paper; Pocono Rabbit Farm and Laboratory Custom Project 35585 Rabbit polyclonal anti-M.HpaII Larsen et al.34 N/A Rabbit polyclonal anti-PEX5 Romano et al.35 N/A Rabbit polyclonal anti-PNKP Stinson et al.17 N/A Rabbit polyclonal anti-FAF1 Kochenova et al.36 N/A Rabbit polyclonal anti-FAF1 Abcam Cat# ab202298 Rabbit polyclonal anti-UBXN7 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Heubes et al.37 N/A Rabbit polyclonal anti-UFD1 Heubes et al.37 N/A Rabbit polyclonal anti-AND1 Antigen: xlAND1 846-1127 This paper; Pocono Rabbit Farm and Laboratory Custom Projects 35808 and 35809 Rabbit polyclonal anti-TRAIP-C Dewar et al.38 N/A Rabbit polyclonal anti-TRAIP-I Antigen: xlTRAIP 336-351 This paper; Vivitide Custom Project 4961 Rabbit polyclonal anti-LacI Antigen: LacI-Bio This paper; BioGenes GmbH Custom Projects 28548 and 28549 Rabbit polyclonal anti-p97 Heubes et al.37 N/A Rabbit polyclonal anti-H3 Cell Signaling Technology Cat# 9715; RRID: AB_331563 Mouse monoclonal anti-Ub Santa Cruz Biotechnology Cat# sc-8017; RRID: AB_628423 Rabbit monoclonal anti-K48 polyubiquitin Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Rabbit monoclonal anti-K63 polyubiquitin Abcam Cat# ab179434; RRID:AB_2895239 Goat Anti-Rabbit IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 111-035-003; RRID: AB_2313567 Mouse Anti-Rabbit IgG (L), HRPconjugated Jackson ImmunoResearch Cat# 211-032-171; RRID: AB_2339149 Rabbit Anti-Mouse IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 315-035-003; RRID: AB_2340061 Mouse Anti-Rabbit IgG (H+L), unconjugated Jackson ImmunoResearch Cat# 211-005-109; RRID: AB_2339147 Streptavidin (HRP) Abcam Cat# ab7403 Bacterial and virus strains E. coli OverExpressTM C41(DE3) Chemically Competent Cells Sigma Cat# CMC0017 Chemicals, peptides, and recombinant proteins NEBuilder HiFi DNA Assembly Master Mix New England Biolabs Cat# E2621L IPTG Sigma Cat# I5502 DTT Bio-Rad Cat# 1610611 cOmpleteTM EDTA-free Protease Inhibitor Cocktail Roche Cat# 11873580001 (Continued on next page) Cell Reports 42, 112125, February 28, 2023 15

Techniques: Ubiquitin Proteomics, Methylation, In Vitro, Recombinant, Concentration Assay, Variant Assay

Journal: Cell reports

Article Title: TRIM21-dependent target protein ubiquitination mediates cell-free Trim-Away.

doi: 10.1016/j.celrep.2023.112125

Figure Lengend Snippet: Figure 7. TRIM21 is ubiquitinated on lysine residues during cell-free Trim-Away (A) Schematic comparing potential TRIM21 ubiquitination sites (pink): N terminus (left panel) and/or lysine residues (right panel). (B) Cell-free Trim-Away assay targeting endogenous TFIIS in HSS. Comparison of the following TRIM21R-R-PS constructs: (1) wild-type (WT), which can be modified on both the N terminus and on lysines; (2) methylated TRIM21 (meWT), which cannot be ubiquitinated on any primary amine; (3) lysine-less TRIM21 (DK) containing a ubiquitinatable N terminus; and (4) a ubiquitin (K63R)-TRIM21 fusion protein (UbK63R-WT), which can be ubiquitinated on lysines only. See Figure S6 for additional schematics. (C) Reconstitution of antibody-dependent target ubiquitination by TRIM21. Recombinant TFIIS was incubated with ATP, ubiquitin, E1, indicated E2 enzymes, and TRIM21R-R-PS in the absence or presence of TFIIS antibody. (D) Proposed Trim-Away model. Upon TRIM21 recruitment to antibody-bound targets, E3 ligase activation leads to direct polyubiquitination of all components on lysines. Substrate modification with heterotypic K48/K63-mixed or -branched polyubiquitin chains mediates its proteasomal degradation. See also Figures S6 and S7.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit monoclonal anti-TRIM21 Cell Signaling Technology Cat# 92043; RRID:AB_2800177 Rabbit polyclonal anti-TFIIS Antigen: xlTFIIS full-length This paper; Pocono Rabbit Farm and Laboratory Custom Project 35585 Rabbit polyclonal anti-M.HpaII Larsen et al.34 N/A Rabbit polyclonal anti-PEX5 Romano et al.35 N/A Rabbit polyclonal anti-PNKP Stinson et al.17 N/A Rabbit polyclonal anti-FAF1 Kochenova et al.36 N/A Rabbit polyclonal anti-FAF1 Abcam Cat# ab202298 Rabbit polyclonal anti-UBXN7 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Kochenova et al.36 N/A Rabbit polyclonal anti-NPL4 Heubes et al.37 N/A Rabbit polyclonal anti-UFD1 Heubes et al.37 N/A Rabbit polyclonal anti-AND1 Antigen: xlAND1 846-1127 This paper; Pocono Rabbit Farm and Laboratory Custom Projects 35808 and 35809 Rabbit polyclonal anti-TRAIP-C Dewar et al.38 N/A Rabbit polyclonal anti-TRAIP-I Antigen: xlTRAIP 336-351 This paper; Vivitide Custom Project 4961 Rabbit polyclonal anti-LacI Antigen: LacI-Bio This paper; BioGenes GmbH Custom Projects 28548 and 28549 Rabbit polyclonal anti-p97 Heubes et al.37 N/A Rabbit polyclonal anti-H3 Cell Signaling Technology Cat# 9715; RRID: AB_331563 Mouse monoclonal anti-Ub Santa Cruz Biotechnology Cat# sc-8017; RRID: AB_628423 Rabbit monoclonal anti-K48 polyubiquitin Cell Signaling Technology Cat# 8081; RRID: AB_10859893 Rabbit monoclonal anti-K63 polyubiquitin Abcam Cat# ab179434; RRID:AB_2895239 Goat Anti-Rabbit IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 111-035-003; RRID: AB_2313567 Mouse Anti-Rabbit IgG (L), HRPconjugated Jackson ImmunoResearch Cat# 211-032-171; RRID: AB_2339149 Rabbit Anti-Mouse IgG (H+L), HRPconjugated Jackson ImmunoResearch Cat# 315-035-003; RRID: AB_2340061 Mouse Anti-Rabbit IgG (H+L), unconjugated Jackson ImmunoResearch Cat# 211-005-109; RRID: AB_2339147 Streptavidin (HRP) Abcam Cat# ab7403 Bacterial and virus strains E. coli OverExpressTM C41(DE3) Chemically Competent Cells Sigma Cat# CMC0017 Chemicals, peptides, and recombinant proteins NEBuilder HiFi DNA Assembly Master Mix New England Biolabs Cat# E2621L IPTG Sigma Cat# I5502 DTT Bio-Rad Cat# 1610611 cOmpleteTM EDTA-free Protease Inhibitor Cocktail Roche Cat# 11873580001 (Continued on next page) Cell Reports 42, 112125, February 28, 2023 15

Techniques: Ubiquitin Proteomics, Comparison, Construct, Methylation, Recombinant, Incubation, Activation Assay

Journal: Arthritis research & therapy

Article Title: Anti-Ro52 positivity is associated with progressive interstitial lung disease in systemic sclerosis-an exploratory study.

doi: 10.1186/s13075-023-03141-4

Figure Lengend Snippet: Fig. 2 Anti-Ro52 positivity predicts decline in vital capacity. Lung function, expressed as percentage of predicted vital capacity (%pVC) over time, declines in anti-Ro52 seropositive systemic sclerosis patients with interstitial lung disease (-2.41%pVC/year; 95% CI: -4.28 – -0.54; p = 0.013). Prediction model using mixed model regression equation including anti-Ro52, anti-Scl-70, disease subtype, and immunomodulator use, with time-interaction terms, and sex; 95% confidence intervals indicated by shaded ribbons

Article Snippet: Monospecific and dual staining was performed with rabbit anti-Trim21 (anti-Ro52) monoclonal antibody (Cell signaling technology, Massachusetts, USA, cat no #92,043, dilution 1:500) and rabbit anti-CD206 polyclonal antibody (Abcam, Cambridge, UK, cat no. ab64693, dilution 1:4000).

Techniques:

Journal: Arthritis research & therapy

Article Title: Anti-Ro52 positivity is associated with progressive interstitial lung disease in systemic sclerosis-an exploratory study.

doi: 10.1186/s13075-023-03141-4

Figure Lengend Snippet: Fig. 3 Anti-Ro52 concentration predicts decline in vital capacity. Lung function, expressed as percentage of predicted vital capacity (%pVC), decreases at a faster rate in patients with systemic sclerosis-associated interstitial lung disease and high anti-Ro52 levels (-0.03%pVC/year; 95% CI: -0.05 – -0.02; p = < 0.001). Prediction model using mixed model regression equation including anti-Ro52 concentration, anti-Scl-70, disease subtype, and immunomodulator use, with time-interaction terms, and sex; 95% confidence intervals indicated by shaded ribbons

Article Snippet: Monospecific and dual staining was performed with rabbit anti-Trim21 (anti-Ro52) monoclonal antibody (Cell signaling technology, Massachusetts, USA, cat no #92,043, dilution 1:500) and rabbit anti-CD206 polyclonal antibody (Abcam, Cambridge, UK, cat no. ab64693, dilution 1:4000).

Techniques: Concentration Assay

Journal: Arthritis research & therapy

Article Title: Anti-Ro52 positivity is associated with progressive interstitial lung disease in systemic sclerosis-an exploratory study.

doi: 10.1186/s13075-023-03141-4

Figure Lengend Snippet: Fig. 5 Ro52 present in peripheral lung tissue from SSc patients IHC from peripheral lung tissue from SSc patient 1 (A-D and SSc patient 2 (E–F) showing cellular localization of Ro52 (brown) in peripheral lung tissue (A, C, E). Co-staining with anti-CD206 and anti-Ro52 showed co-localization in alveolar M2 macrophages (B, D, F). Scale bar = 100 µm

Article Snippet: Monospecific and dual staining was performed with rabbit anti-Trim21 (anti-Ro52) monoclonal antibody (Cell signaling technology, Massachusetts, USA, cat no #92,043, dilution 1:500) and rabbit anti-CD206 polyclonal antibody (Abcam, Cambridge, UK, cat no. ab64693, dilution 1:4000).

Techniques: Staining

Journal: Arthritis research & therapy

Article Title: Anti-Ro52 positivity is associated with progressive interstitial lung disease in systemic sclerosis-an exploratory study.

doi: 10.1186/s13075-023-03141-4

Figure Lengend Snippet: Fig. 4 Ro52 present in peripheral lung tissue from healthy organ donors IHC from healthy peripheral lung tissue in donor 1 (A-D) and donor 2 (E–H) showing cellular localization of Ro52 (brown) in peripheral lung tissue (A, E) with distinct expression on alveolar macrophages (CD206; red; B, F). Co-staining with anti-CD206 and anti-Ro52 showed co-localization in alveolar M2 macrophages (C, D, G, H). Scale bar = 20 µm (A-C, E–G), 10 µm (D, H). Representative images from two healthy donors

Article Snippet: Monospecific and dual staining was performed with rabbit anti-Trim21 (anti-Ro52) monoclonal antibody (Cell signaling technology, Massachusetts, USA, cat no #92,043, dilution 1:500) and rabbit anti-CD206 polyclonal antibody (Abcam, Cambridge, UK, cat no. ab64693, dilution 1:4000).

Techniques: Expressing, Staining