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α tgn46 sheep antibody (Bio-Rad)

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Bio-Rad α tgn46 sheep antibody
α Tgn46 Sheep Antibody, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 96/100, based on 752 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/%CE%B1+tgn46+sheep+antibody/Sheep+anti+Human+TGN46/pm41096722-265-12-16
Average 96 stars, based on 752 article reviews

α tgn46 sheep antibody - by Bioz Stars, 2026-08

96/100 stars

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A, B Immunoblots showing results of (A) immunoprecipitation experiments from GFP‐HEATR5B and GFP HEK293 cells and (B) in vitro pull‐downs with recombinant, bead‐associated GFP‐tagged human HEATR5B or GFP protein incubated with purified dynein tail and/or dynactin complexes. G‐HR5B, GFP‐HEATR5B; G, GFP; IP, immunoprecipitate; IB, protein probed by immunoblotting; MW, molecular weight of protein standards (theoretical MW is shown for DYNC1H1 as standards of a similar molecular weight were not available); DT, dynein tail complex; DCTN, dynactin complex. In (A), GAPDH is a negative control. Source data are available online for this figure.

Journal: The EMBO Journal

Article Title: HEATR5B associates with dynein‐dynactin and promotes motility of AP1 ‐bound endosomal membranes

doi: 10.15252/embj.2023114473

Figure Lengend Snippet: A, B Immunoblots showing results of (A) immunoprecipitation experiments from GFP‐HEATR5B and GFP HEK293 cells and (B) in vitro pull‐downs with recombinant, bead‐associated GFP‐tagged human HEATR5B or GFP protein incubated with purified dynein tail and/or dynactin complexes. G‐HR5B, GFP‐HEATR5B; G, GFP; IP, immunoprecipitate; IB, protein probed by immunoblotting; MW, molecular weight of protein standards (theoretical MW is shown for DYNC1H1 as standards of a similar molecular weight were not available); DT, dynein tail complex; DCTN, dynactin complex. In (A), GAPDH is a negative control. Source data are available online for this figure.

Article Snippet: The following antibodies were used for immunoblotting (IB) and immunofluorescence (IF) of human cell extracts or cells, respectively (working dilutions in parentheses): α‐GFP chicken antibody (Abcam 13970; IB, 1:5,000; IF, 1:200), α‐DYNC1H1 rabbit antibody (Proteintech 12345‐1‐AP; IB, 1:100; IF, 1:100), α‐DCTN1 mouse antibody (BD Transduction Laboratories 610474; IB, 1:5,000; IF, 1:50), α‐AFTPH rabbit antibody (Invitrogen PA5‐57104; IB, 1:500), α‐γ‐SYNRG (AP1GBP1) rabbit antibody (Novusbio NBP1‐90145; IB, 1:200), α‐AP1γ (F‐10) mouse antibody (Santa Cruz Sc‐398867; IB, 1:100), α‐AP1γ mouse antibody (Sigma A4200, clone 100.3; IF, 1:400), α‐EEA1 mouse antibody (BD Transduction laboratories 610457; IF, 1:200), α‐LAMP1 mouse antibody (Abcam 25630; IF, 1:200), α‐GAPDH rabbit antibody (Sigma G9545; IB, 1:4,000), α‐HEATR5B rabbit antibody (Hirst et al , ; provided by J. Hirst and M. Robinson [Cambridge Institute for Medical Research, UK]; IB, 1:250), α‐TGN46 sheep antibody (Bio‐Rad AHP500; IF, 1:200), α‐RAB11A rabbit monoclonal antibody (Abcam 128913; IF, 1:50), α‐β‐actin mouse antibody (Gen Tex GTX26276; IB, 1:20,000), α‐α‐Tubulin mouse antibody (Santa Cruz; Sc‐32293 clone DM1A; IF, 1:500), and α‐Transferrin receptor rabbit antibody (Abcam ab84036; IF, 1:200).

Techniques: Western Blot, Immunoprecipitation, In Vitro, Recombinant, Incubation, Purification, Molecular Weight, Negative Control

A Representative confocal images of immunostained HeLa cells showing localisation of GFP‐HEATR5B (HR5B) (GFP signal amplified with GFP antibodies) and AP1γ. Yellow arrows, position of TGN‐associated AP1γ (which rarely co‐localises with GFP‐HR5B); dashed box shows area containing multiple co‐localisation events that is magnified in right‐hand images. In this and other figures, DAPI is used to stain DNA. B Quantification of percentage of GFP‐HR5B puncta that co‐localise with indicated proteins in fixed cells. C Quantification of percentage of GFP‐HR5B puncta that co‐localise with AP1σ1‐RFP in live HeLa cells. D Example kymograph (time‐distance plot) of a long‐range co‐transport event of GFP‐HR5B and AP1σ1‐RFP. E Quantification of percentage of GFP‐HR5B puncta that co‐localise with DsRed‐RAB11A in live HeLa cells. F Example kymograph of a long‐range co‐transport event of GFP‐HR5B and DsRed‐RAB11A. G, H Representative confocal images (G) and quantification (H) of spread of the indicated protein signals away from the perinuclear region in immunostained U2OS cells ± DYNC1H1 siRNA. Data information: In (A and G), white circles indicate merge of magnified images. In (D and F), d = distance and t = time. Scale bars: (A) 10 μm; (A) insets, 2 μm; (D) 20 μm and 20 s; (F) 10 μm and 10 s; (G) 10 μm; (G) insets 2.5 μm. In (B, C, E and H), number of cells analysed is shown above columns. In (B, C, and E), circles indicate values from individual cells, with columns and error bars representing mean ± SD. In (H), boxes show interquartile range (25 th –75 th percentile of values) and horizontal line is the median; statistical significance was evaluated with an unpaired two‐tailed t ‐test: **** P < 0.0001. Source data are available online for this figure.

Journal: The EMBO Journal

Article Title: HEATR5B associates with dynein‐dynactin and promotes motility of AP1 ‐bound endosomal membranes

doi: 10.15252/embj.2023114473

Figure Lengend Snippet: A Representative confocal images of immunostained HeLa cells showing localisation of GFP‐HEATR5B (HR5B) (GFP signal amplified with GFP antibodies) and AP1γ. Yellow arrows, position of TGN‐associated AP1γ (which rarely co‐localises with GFP‐HR5B); dashed box shows area containing multiple co‐localisation events that is magnified in right‐hand images. In this and other figures, DAPI is used to stain DNA. B Quantification of percentage of GFP‐HR5B puncta that co‐localise with indicated proteins in fixed cells. C Quantification of percentage of GFP‐HR5B puncta that co‐localise with AP1σ1‐RFP in live HeLa cells. D Example kymograph (time‐distance plot) of a long‐range co‐transport event of GFP‐HR5B and AP1σ1‐RFP. E Quantification of percentage of GFP‐HR5B puncta that co‐localise with DsRed‐RAB11A in live HeLa cells. F Example kymograph of a long‐range co‐transport event of GFP‐HR5B and DsRed‐RAB11A. G, H Representative confocal images (G) and quantification (H) of spread of the indicated protein signals away from the perinuclear region in immunostained U2OS cells ± DYNC1H1 siRNA. Data information: In (A and G), white circles indicate merge of magnified images. In (D and F), d = distance and t = time. Scale bars: (A) 10 μm; (A) insets, 2 μm; (D) 20 μm and 20 s; (F) 10 μm and 10 s; (G) 10 μm; (G) insets 2.5 μm. In (B, C, E and H), number of cells analysed is shown above columns. In (B, C, and E), circles indicate values from individual cells, with columns and error bars representing mean ± SD. In (H), boxes show interquartile range (25 th –75 th percentile of values) and horizontal line is the median; statistical significance was evaluated with an unpaired two‐tailed t ‐test: **** P < 0.0001. Source data are available online for this figure.

Techniques: Amplification, Staining, Two Tailed Test

A, B Representative confocal images of wild‐type (A) and stable GFP‐HEATR5B (B) HeLa cells stained with antibodies to the indicated proteins (GFP signal in panel (B) was amplified with GFP antibodies). Dashed box shows area magnified in right‐hand images. Panel (A) shows that AP1γ is clustered at the periphery of the TGN. Panel (B) shows that association of GFP‐HEATR5B (HR5B) with TGN46, EEA1 and LAMP1 is rarely observed. Scale bars: main panels, 10 μm; insets, 2.5 μm. White circles indicate merge of magnified images.

Journal: The EMBO Journal

Article Title: HEATR5B associates with dynein‐dynactin and promotes motility of AP1 ‐bound endosomal membranes

doi: 10.15252/embj.2023114473

Figure Lengend Snippet: A, B Representative confocal images of wild‐type (A) and stable GFP‐HEATR5B (B) HeLa cells stained with antibodies to the indicated proteins (GFP signal in panel (B) was amplified with GFP antibodies). Dashed box shows area magnified in right‐hand images. Panel (A) shows that AP1γ is clustered at the periphery of the TGN. Panel (B) shows that association of GFP‐HEATR5B (HR5B) with TGN46, EEA1 and LAMP1 is rarely observed. Scale bars: main panels, 10 μm; insets, 2.5 μm. White circles indicate merge of magnified images.

Techniques: Staining, Amplification

A Representative confocal images of AP1γ and RAB11A in immunostained wild‐type (control) and HEATR5B mutant ( HR5B KO) U2OS cells. Dashed box shows region magnified in right‐hand images. Blue arrowheads show diffuse AP1γ signal in the cytoplasm of mutant cells. Yellow arrowheads in insets show examples of AP1γ association with RAB11A. White circles indicate merge of magnified images. B, C Quantification of number (B) and mean total intensity (C) of AP1γ puncta in control (Ctrl) and HR5B KO U2OS cells (a.u., arbitrary units). D Left, immunoblot images showing levels of HR5B and AP1γ in control and HR5B KO U2OS cells (β‐actin, loading control). Right, quantification of AP1γ signal (normalised to β‐actin signal). E Quantification of spread of total AP1γ signal away from the perinuclear region in control and HR5B KO U2OS cells. Both punctate and diffuse signals were quantified. F Quantification of weighted mean MSD of AP1σ1‐RFP puncta in control and HR5B KO U2OS cells. G, H Representative confocal images showing localisation of RAB11A (G and H), AP1γ (G) and DCTN1 (H) in cells that strongly express GFP‐HR5B or a GFP control. Arrowheads show perinuclear clustering of signals in GFP‐HR5B cells. GFP signals were not amplified with antibodies. I Quantification of spread of RAB11A signal away from perinuclear region in GFP or GFP‐HR5B overexpressing U2OS cells. Data information: Scale bars: (A) 10 μm; (A) insets 2.5 μm; (G and H) 5 μm. In (B, C, and E), boxes show interquartile range (25 th –75 th percentile of values) and horizontal line is the median. In (I), circles indicate values from individual cells; columns and error bars represent mean ± SD. Number of cells (in B, C, E and I) or independent experiments (D) analysed is shown above columns. In (F), n = number of particles (from 31 control and 17 KO U2OS cells). Statistical significance was evaluated with an unpaired two‐tailed t ‐test: **** P < 0.0001; *** P < 0.001; ** P < 0.01. Source data are available online for this figure.

Journal: The EMBO Journal

Article Title: HEATR5B associates with dynein‐dynactin and promotes motility of AP1 ‐bound endosomal membranes

doi: 10.15252/embj.2023114473

Figure Lengend Snippet: A Representative confocal images of AP1γ and RAB11A in immunostained wild‐type (control) and HEATR5B mutant ( HR5B KO) U2OS cells. Dashed box shows region magnified in right‐hand images. Blue arrowheads show diffuse AP1γ signal in the cytoplasm of mutant cells. Yellow arrowheads in insets show examples of AP1γ association with RAB11A. White circles indicate merge of magnified images. B, C Quantification of number (B) and mean total intensity (C) of AP1γ puncta in control (Ctrl) and HR5B KO U2OS cells (a.u., arbitrary units). D Left, immunoblot images showing levels of HR5B and AP1γ in control and HR5B KO U2OS cells (β‐actin, loading control). Right, quantification of AP1γ signal (normalised to β‐actin signal). E Quantification of spread of total AP1γ signal away from the perinuclear region in control and HR5B KO U2OS cells. Both punctate and diffuse signals were quantified. F Quantification of weighted mean MSD of AP1σ1‐RFP puncta in control and HR5B KO U2OS cells. G, H Representative confocal images showing localisation of RAB11A (G and H), AP1γ (G) and DCTN1 (H) in cells that strongly express GFP‐HR5B or a GFP control. Arrowheads show perinuclear clustering of signals in GFP‐HR5B cells. GFP signals were not amplified with antibodies. I Quantification of spread of RAB11A signal away from perinuclear region in GFP or GFP‐HR5B overexpressing U2OS cells. Data information: Scale bars: (A) 10 μm; (A) insets 2.5 μm; (G and H) 5 μm. In (B, C, and E), boxes show interquartile range (25 th –75 th percentile of values) and horizontal line is the median. In (I), circles indicate values from individual cells; columns and error bars represent mean ± SD. Number of cells (in B, C, E and I) or independent experiments (D) analysed is shown above columns. In (F), n = number of particles (from 31 control and 17 KO U2OS cells). Statistical significance was evaluated with an unpaired two‐tailed t ‐test: **** P < 0.0001; *** P < 0.001; ** P < 0.01. Source data are available online for this figure.

Techniques: Control, Mutagenesis, Western Blot, Amplification, Two Tailed Test

( A ) Immunofluorescent staining of different Golgi marker, the matrix proteins GM130 (green), the cis -Golgi protein Giantin (red) and the trans -Golgi network protein TGN46 (magenta) in representative control (top) and MYO1C depleted (bottom) hTertRPE-1 cells in classical, unconstrained culture conditions. The nucleus is stained with Dapi (blue). Quantification of the 2D projected Golgi area in control conditions (n=145) and upon MYO1C depletion (n=165). Error bars represent the standard deviation of three independent experiments. *** indicates P-value < 1×10 −4 in a Student T-test. ( C ) Fluorescent images of representative hTertRPE-1 cells expressing GFP or GFP-MYO1C (green) and stained for the Golgi apparatus (red, GM130). ( D ) Quantification of the 2D projected Golgi area (stained by GM130) of non transfected cells and cells as in C. Error bars represent the standard deviation of three independent experiments with n > 40 cells of each condition. * indicates P-value < 1×10 −2 in a Student T-test on averages of three independent experiments ( E ) Electron microscopy images of representative intracellular areas of unconstrained hTertRPE-1 cells containing the Golgi apparatus in control conditions and upon MYO1C depletion. Scale bar: 1 µ m.

Journal: bioRxiv

Article Title: MYO1C facilitates arrival at the Golgi apparatus through stabilization of branched actin

doi: 10.1101/409110

Figure Lengend Snippet: ( A ) Immunofluorescent staining of different Golgi marker, the matrix proteins GM130 (green), the cis -Golgi protein Giantin (red) and the trans -Golgi network protein TGN46 (magenta) in representative control (top) and MYO1C depleted (bottom) hTertRPE-1 cells in classical, unconstrained culture conditions. The nucleus is stained with Dapi (blue). Quantification of the 2D projected Golgi area in control conditions (n=145) and upon MYO1C depletion (n=165). Error bars represent the standard deviation of three independent experiments. *** indicates P-value < 1×10 −4 in a Student T-test. ( C ) Fluorescent images of representative hTertRPE-1 cells expressing GFP or GFP-MYO1C (green) and stained for the Golgi apparatus (red, GM130). ( D ) Quantification of the 2D projected Golgi area (stained by GM130) of non transfected cells and cells as in C. Error bars represent the standard deviation of three independent experiments with n > 40 cells of each condition. * indicates P-value < 1×10 −2 in a Student T-test on averages of three independent experiments ( E ) Electron microscopy images of representative intracellular areas of unconstrained hTertRPE-1 cells containing the Golgi apparatus in control conditions and upon MYO1C depletion. Scale bar: 1 µ m.

Article Snippet: We used the following mouse monoclonal antibodies α-GM130 (BD Biosciences cat: 610823), α-MYO1C (Santa Cruz sc-136544) and α-actin (Sigma, A-2228), rabbit polyclonal antibodies α-Lamp1 (Sigma, L1418 or Abcam, Ab24170), human monoclonal antibody (F2C-hFc) α-tubulin and α-Giantin (Recombinant Protein and Antibody Platform of the Institut Curie) and sheep polyclonal antibody α-TGN46 (Bio-Rad, AHP500).

Techniques: Staining, Marker, Standard Deviation, Expressing, Transfection, Electron Microscopy

( A ) Fluorescent images of a representative, unconstrained hTertRPE-1 cell stained with MYO1C antibody to visualize the endogenous protein (green), TGN46 antibody to visualize the Golgi apparatus (red) and Dapi to visualize the nucleus (blue). ( B, C ) Fluorescent images of representative, micropatterned hTertRPE-1 cells expressing GFP-MYO1C (green) and stained for the Golgi apparatus either with TGN46 or Giantin antibody (red) and the nucleus with Dapi (blue). ( D ) Fluorescent images of a time-lapse acquisition of unconstrained hTertRPE-1 cells expressing GFP-MYO1C (green) and mCherry-Rab6 (red). Scale bar: 10 µm.

Journal: bioRxiv

Article Title: MYO1C facilitates arrival at the Golgi apparatus through stabilization of branched actin

doi: 10.1101/409110

Figure Lengend Snippet: ( A ) Fluorescent images of a representative, unconstrained hTertRPE-1 cell stained with MYO1C antibody to visualize the endogenous protein (green), TGN46 antibody to visualize the Golgi apparatus (red) and Dapi to visualize the nucleus (blue). ( B, C ) Fluorescent images of representative, micropatterned hTertRPE-1 cells expressing GFP-MYO1C (green) and stained for the Golgi apparatus either with TGN46 or Giantin antibody (red) and the nucleus with Dapi (blue). ( D ) Fluorescent images of a time-lapse acquisition of unconstrained hTertRPE-1 cells expressing GFP-MYO1C (green) and mCherry-Rab6 (red). Scale bar: 10 µm.

Techniques: Staining, Expressing

Journal: bioRxiv

Article Title: MYO1C facilitates arrival at the Golgi apparatus through stabilization of branched actin

doi: 10.1101/409110

Figure Lengend Snippet: ( A ) Fluorescent images of a representative, unconstrained hTertRPE-1 cell stained with MYO1C antibody to visualize the endogenous protein (green), TGN46 antibody to visualize the Golgi apparatus (magenta) and phalloidin to visualize the actin cytoskeleton (red). The nucleus is stained with Dapi (blue). ( B ) Fluorescent images of representative, micropatterned hTertRPE-1 cells expressing GFP-MYO1C (green) and stained with phalloidin to visualize F-actin (red) and Dapi to visualize the nucleus (blue). ( C ) Domain structure of MYO1C, the mutant form MYO1CΔABL and the truncated Tail domain. ( D ) Fluorescent images of representative hTertRPE-1 cells expressing GFP-MYO1C, GFP-MYO1CΔABL or GFP-Tail (green), stained for the Golgi apparatus (magenta, GM130) and F-actin with phalloidin (red). ( E ) Quantification of GFP-positive spots at the Golgi area as in D for GFP (n=24), GFP-MYO1C (n=49), GFP-MYO1CΔABL (n=56) or GFP-Tail (n=34). % of cells is shown that contain 0 (light grey), 1-10 (dark grey) or more than 10 (black) Golgi-associated spots per cell. Note that control (GFP-expressing) cells never contained more than 1 Golgi-associated spot per cell.

Techniques: Staining, Expressing, Mutagenesis

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