anti collagen iv  (Abcam)

 
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    Name:
    Anti Collagen IV antibody C IV 22
    Description:

    Catalog Number:
    ab15633
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    Structured Review

    Abcam anti collagen iv

    https://www.bioz.com/result/anti collagen iv/product/Abcam
    Average 98 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    anti collagen iv - by Bioz Stars, 2020-07
    98/100 stars

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    Produced:

    Article Title: The Plasminogen Receptor, Plg-RKT, is Essential for Mammary Lobuloalveolar Development and Lactation
    Article Snippet: .. Primary antibodies used for immunohistochemical staining of paraffin embedded tissue were: anti-collagen IV and anti-α-smooth muscle actin (Abcam), anti-laminin 1 (R & D Systems), anti-mouse β1 integrin (Chemicon), anti-mouse F4/80 (Biolegend),, anti-human fibrinogen (Dako), and anti-mouse Plg-RKT , produced in our laboratory [ ]. .. For immunohistochemistry of frozen sections, anti-Ser28 -phosphorylated histone H3 was from Sigma.

    Western Blot:

    Article Title: High glucose induces platelet‐derived growth factor‐C via carbohydrate response element‐binding protein in glomerular mesangial cells. High glucose induces platelet‐derived growth factor‐C via carbohydrate response element‐binding protein in glomerular mesangial cells
    Article Snippet: .. For western blots, the following primary antibodies were used: anti‐PDGF‐C (Abcam, Cambridge, UK), anti‐Collagen IV (Abcam), anti‐Collagen VI (Abcam), and anti‐b‐actin (Santa Cruz, Dallas, TX) and they were diluted 1:1000. .. Primary antibodies were incubated in Tris‐buffered saline with 1% milk.

    Incubation:

    Article Title: Long non-coding RNA MEG3 impacts diabetic nephropathy progression through sponging miR-145
    Article Snippet: .. The membrane was blocked with 5% nonfat milk in PBS and then incubated with anti-Collagen IV, anti-Fibronectin and anti-Bcl-2 in PBS overnight, followed by an incubation of horseradish peroxidase conjugated secondary antibody (Abcam, USA) for 2 h at room temperature. .. Blots were processed with an enhanced chemiluminescence kit (Santa Cruz Biotechnology), and exposed to the film.

    Article Title: Fabrication of Cardiac Patch with Decellularized Porcine Myocardial Scaffold and Bone Marrow Mononuclear Cells
    Article Snippet: .. For immunofluorescent staining, after rehydration and antigen retrieval with 0.05% Trypsin for 10 minutes at 37°C, sections were washed and incubated at 4 °C for overnight with sarcomeric α-actinin (sigma), myosin heavy chain (GenWay), cardiac troponin T, von Willebrand factor (vWF), and Collagen IV (abcam) primary antibodies. .. After the primary antibody, Cy3 AffiniPure Goat Anti-Mouse IgG (H+L), Cy5 AffiniPure Donkey Anti-Mouse IgM, Alexa Fluor secondary antibodies (JacksonImmuno Research), Goat Anti-Mouse IgG, and Goat Anti-Rabbit IgG secondary antibodies (abcam) were incubated at 37°C for 30 minutes to obtain fluorescent colors.

    Article Title: Nanoengineered, cell-derived extracellular matrix influences ECM-related gene expression of mesenchymal stem cells
    Article Snippet: .. Then, cells were permeabilized in 1% BSA and 0.3% Triton X-100 in PBS for 10 min. After washing the permeabilizing solution, cells were incubated with the targeted specific primary antibodies overnight at 4 °C: anti-fibronectin (1:200 dilution, Santa Cruz, Deleware, USA), anti-collagen type I (1:200 dilution, Abcam, Cambridge, UK), anti-collagen type II (1:200 dilution, Abcam, Cambridge, UK), anti-collagen IV (1:200 dilution, Abcam, Cambridge, UK) and anti-laminin (1:100 dilution, Sigma, St. Louis, USA). .. After washing cells 3 times with DPBS, cells were incubated with the secondary antibody (1:300 dilution, Goat anti-rabbit IgG –TRITC) (Sigma, St. Louis, USA) for 1 h at room temperature.

    Immunohistochemistry:

    Article Title: Inflammatory Smooth Muscle Cells Induce Endothelial Cell Alterations to Influence Cerebral Aneurysm Progression via Regulation of Integrin and VEGF Expression
    Article Snippet: .. Anti-von Willebrand Factor (vWF, Abcam, Cambridge, MA, USA), anti-alpha smooth muscle actin (αSMA, Dako, Carpinteria, CA, USA), anti-Angiotensin II Type 1 Receptor (AT1, Novus Biologicals, Cambridge, UK), anti-MMP (matrix metalloproteinase)-1 (Millipore, Billerica, MA,USA), anti-MMP-3 (Millipore), anti-collagen IV (Abcam), and anti-collagen VI (Abcam) primary antibodies were used for IHC. .. 3,3′-diaminobenzidine (DAB) plus chromogen (Thermo Fisher Scientific, Waltham, MA, USA) was used for substrate visualization, according to the manufacturer’s protocol.

    Article Title: Therapeutic Potential of Nrf2 Activators in Streptozotocin-Induced Diabetic Nephropathy
    Article Snippet: .. Antibodies for immunoblot and immunohistochemical (IHC) analysis included anti-7,8-dihydro-8-oxo-2’-deoxyguanosine (8-oxo-dG) (Trevigen, Gaithersburg, MD); anti-fibronectin (FN) (Calbiochem, San Diego, CA); anti-collagen IV (Abcam, San Francisco, CA); anti-Ki67 (Vector Laboratories, Burlingame, CA); and anti-Nrf2, NAD(P)H dehydrogenase, quinone 1 (NQO1), γ-glutamylcysteine synthetase (γ-GCS), p21, TGF-β1, and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA). ..

    Article Title: The Plasminogen Receptor, Plg-RKT, is Essential for Mammary Lobuloalveolar Development and Lactation
    Article Snippet: .. Primary antibodies used for immunohistochemical staining of paraffin embedded tissue were: anti-collagen IV and anti-α-smooth muscle actin (Abcam), anti-laminin 1 (R & D Systems), anti-mouse β1 integrin (Chemicon), anti-mouse F4/80 (Biolegend),, anti-human fibrinogen (Dako), and anti-mouse Plg-RKT , produced in our laboratory [ ]. .. For immunohistochemistry of frozen sections, anti-Ser28 -phosphorylated histone H3 was from Sigma.

    Staining:

    Article Title: Fabrication of Cardiac Patch with Decellularized Porcine Myocardial Scaffold and Bone Marrow Mononuclear Cells
    Article Snippet: .. For immunofluorescent staining, after rehydration and antigen retrieval with 0.05% Trypsin for 10 minutes at 37°C, sections were washed and incubated at 4 °C for overnight with sarcomeric α-actinin (sigma), myosin heavy chain (GenWay), cardiac troponin T, von Willebrand factor (vWF), and Collagen IV (abcam) primary antibodies. .. After the primary antibody, Cy3 AffiniPure Goat Anti-Mouse IgG (H+L), Cy5 AffiniPure Donkey Anti-Mouse IgM, Alexa Fluor secondary antibodies (JacksonImmuno Research), Goat Anti-Mouse IgG, and Goat Anti-Rabbit IgG secondary antibodies (abcam) were incubated at 37°C for 30 minutes to obtain fluorescent colors.

    Article Title: The Plasminogen Receptor, Plg-RKT, is Essential for Mammary Lobuloalveolar Development and Lactation
    Article Snippet: .. Primary antibodies used for immunohistochemical staining of paraffin embedded tissue were: anti-collagen IV and anti-α-smooth muscle actin (Abcam), anti-laminin 1 (R & D Systems), anti-mouse β1 integrin (Chemicon), anti-mouse F4/80 (Biolegend),, anti-human fibrinogen (Dako), and anti-mouse Plg-RKT , produced in our laboratory [ ]. .. For immunohistochemistry of frozen sections, anti-Ser28 -phosphorylated histone H3 was from Sigma.

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  • 90
    Abcam type iv collagens anti type iv collagen rabbit polyclonal antibody
    Screening of antibodies by immunoprecipitation and western blotting . Three micrograms of each antibody designated in the figure was incubated with the supernatant of HLF cell cultured medium and the antibody was precipitated with anti-mouse IgG antibodies. The precipitates were subjected to western blotting under nonreducing condition. A <t>polyclonal</t> antibody for human type IV collagen, Ab6586, was used to detect NTH α1(IV) and type IV collagen polypeptides. A band migrating at 180 kDa on the blot is NTH α1(IV). A band migrating at ~500 kDa corresponds to a trimer of three α chains of the type IV collagen molecule, which are connected by interchain disulfide bondings. IP, immunoprecipitation; SUP, supernatant; CTN, control. ( a ) In the first screening, immunoprecipitates of seven prepared antibodies #141, #370, #792, #1283, #495, #702 and #189, were tested for the samples developed on 4.5% SDS-PAGE under nonreducing conditions. Three antibodies, #141, #370 and #1283, were selected for further analysis by sandwich ELISA to distinguish overlapping epitopes. ( b ) In the second screening, only #179 was selected because of its unique binding characteristics, as described in Table 2 .
    Type Iv Collagens Anti Type Iv Collagen Rabbit Polyclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/type iv collagens anti type iv collagen rabbit polyclonal antibody/product/Abcam
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    type iv collagens anti type iv collagen rabbit polyclonal antibody - by Bioz Stars, 2020-07
    90/100 stars
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    89
    Abcam rabbit anti mouse collagen iv
    PECAM and CD99 function past the level of the endothelium in C57BL/6 mice. A : wild-type FVB/n and C57BL/6 mice were pretreated with control IgG, <t>anti-PECAM,</t> or anti-CD99 (3 mg/kg) prior to ear stimulation with croton oil (1% solution, 20 μl/ear) or carrier alone. After 4 h, ear tissue was collected and immunohistochemical staining was performed for PECAM [endothelial cells (ECs), red], myeloid-related protein 14 (neutrophils, green), and <t>collagen</t> <t>IV</t> [basement membrane (BM), purple]. 3-Dimensional confocal images were acquired from each sample and subsequently analyzed. Insets : z -orthogonal view along the indicated line to demonstrate leukocyte arrest in relation to the endothelium and basement membrane at 1.5 times the xy image. Scale bar = 50 μm. B : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in FVB/n mice). C : schematic of quantification of site of leukocyte blockade. Leukocytes were scored as being in 1 of 6 positions: 1 ) luminal, 2 ) apically arrested, 3 ) arrested partway through the endothelium, 4 ) arrested at the level of the basement membrane, 5 ) actively migrating through the basement membrane, or 6 ) fully extravasated. D : quantification of the site of leukocyte arrest in FVB/n mice. E : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in C57BL/6 mice). F : quantification of the site of leukocyte arrest in C57BL/6 mice. G : schematic model of anti-PECAM and anti-CD99 arrest in FVB/n and C57BL/6 mice. One hundred to 200 cells were analyzed per ear. Carrier-alone stimulation resulted in minimal leukocyte recruitment (data not shown). Total leukocytes per field of view, vessel length, and vessel diameter were equivalent for all treatment groups (data not shown). Data are representative of recordings from ≥4 mice per group with ≥8 images quantified per <t>mouse.</t> * P
    Rabbit Anti Mouse Collagen Iv, supplied by Abcam, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti mouse collagen iv/product/Abcam
    Average 89 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    rabbit anti mouse collagen iv - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    94
    Abcam anti mouse cytochrome c
    Rab5 regulates <t>cytochrome</t> c release during H 2 O 2 -induced stress. ( A ) Live-cell imaging of a BAC GFP-Rab5 cell stained with 100 nM Mito-Red before (0 min) and after treatment (60 min) with 250 μM H 2 O 2 at 37°C. Inset images are shown and arrowheads indicate GFP-Rab5 recruitment to MOMP events (right panel). Scale bars, 10 μm. ( B ) HeLa cells over-expressing either GFP or GFP-Rab5 were treated with 250 μM H 2 O 2 at 37°C for 1 to 4 hr. Protein samples from cytosolic fractions were obtained and immunoblotted for cytochrome c and tubulin (as a loading control). ( C ) Densitometric quantification of cytochrome c release in ( B ). Data were collected from three independent experiments. Y-axis corresponds to the normalized ratio intensity of cytochrome c to the tubulin loading control. p-Values based on two-tailed t-tests. 10.7554/eLife.32282.034 Numerical data corresponding to the line traces presented in Figure 7C .
    Anti Mouse Cytochrome C, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mouse cytochrome c/product/Abcam
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mouse cytochrome c - by Bioz Stars, 2020-07
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    Image Search Results


    Screening of antibodies by immunoprecipitation and western blotting . Three micrograms of each antibody designated in the figure was incubated with the supernatant of HLF cell cultured medium and the antibody was precipitated with anti-mouse IgG antibodies. The precipitates were subjected to western blotting under nonreducing condition. A polyclonal antibody for human type IV collagen, Ab6586, was used to detect NTH α1(IV) and type IV collagen polypeptides. A band migrating at 180 kDa on the blot is NTH α1(IV). A band migrating at ~500 kDa corresponds to a trimer of three α chains of the type IV collagen molecule, which are connected by interchain disulfide bondings. IP, immunoprecipitation; SUP, supernatant; CTN, control. ( a ) In the first screening, immunoprecipitates of seven prepared antibodies #141, #370, #792, #1283, #495, #702 and #189, were tested for the samples developed on 4.5% SDS-PAGE under nonreducing conditions. Three antibodies, #141, #370 and #1283, were selected for further analysis by sandwich ELISA to distinguish overlapping epitopes. ( b ) In the second screening, only #179 was selected because of its unique binding characteristics, as described in Table 2 .

    Journal: Biochemistry and Biophysics Reports

    Article Title: Preparation and partial characterization of monoclonal antibodies specific for the nascent non-triple helical form of the type IV collagen alpha 1 chain

    doi: 10.1016/j.bbrep.2016.11.013

    Figure Lengend Snippet: Screening of antibodies by immunoprecipitation and western blotting . Three micrograms of each antibody designated in the figure was incubated with the supernatant of HLF cell cultured medium and the antibody was precipitated with anti-mouse IgG antibodies. The precipitates were subjected to western blotting under nonreducing condition. A polyclonal antibody for human type IV collagen, Ab6586, was used to detect NTH α1(IV) and type IV collagen polypeptides. A band migrating at 180 kDa on the blot is NTH α1(IV). A band migrating at ~500 kDa corresponds to a trimer of three α chains of the type IV collagen molecule, which are connected by interchain disulfide bondings. IP, immunoprecipitation; SUP, supernatant; CTN, control. ( a ) In the first screening, immunoprecipitates of seven prepared antibodies #141, #370, #792, #1283, #495, #702 and #189, were tested for the samples developed on 4.5% SDS-PAGE under nonreducing conditions. Three antibodies, #141, #370 and #1283, were selected for further analysis by sandwich ELISA to distinguish overlapping epitopes. ( b ) In the second screening, only #179 was selected because of its unique binding characteristics, as described in Table 2 .

    Article Snippet: 2.2 Antibodies and type IV collagens Anti-type IV collagen rabbit polyclonal antibody, Ab6586, was purchased from Abcam (Tokyo, Japan).

    Techniques: Immunoprecipitation, Western Blot, Incubation, Cell Culture, SDS Page, Sandwich ELISA, Binding Assay

    Location of type IV collagen and type VII collagen in normal and colorectal cancer tissue from the same patient . Frozen tissue sections were incubated on the glass slide with polyclonal rabbit anti-type IV collagen and monoclonal mouse anti-type VII collagen. Negative controls without primary antibody were run in parallel (not shown). The bar represents 50 μm.

    Journal: BMC Cancer

    Article Title: Collagen mRNA levels changes during colorectal cancer carcinogenesis

    doi: 10.1186/1471-2407-9-136

    Figure Lengend Snippet: Location of type IV collagen and type VII collagen in normal and colorectal cancer tissue from the same patient . Frozen tissue sections were incubated on the glass slide with polyclonal rabbit anti-type IV collagen and monoclonal mouse anti-type VII collagen. Negative controls without primary antibody were run in parallel (not shown). The bar represents 50 μm.

    Article Snippet: The sections were examined by immunofluorescence microscopy for the localization of type IV and type VII collagens using polyclonal rabbit anti type IV collagen (Abcam 6586) and monoclonal mouse anti-type VII collagen (Abcam 6312), respectively.

    Techniques: Incubation

    PECAM and CD99 function past the level of the endothelium in C57BL/6 mice. A : wild-type FVB/n and C57BL/6 mice were pretreated with control IgG, anti-PECAM, or anti-CD99 (3 mg/kg) prior to ear stimulation with croton oil (1% solution, 20 μl/ear) or carrier alone. After 4 h, ear tissue was collected and immunohistochemical staining was performed for PECAM [endothelial cells (ECs), red], myeloid-related protein 14 (neutrophils, green), and collagen IV [basement membrane (BM), purple]. 3-Dimensional confocal images were acquired from each sample and subsequently analyzed. Insets : z -orthogonal view along the indicated line to demonstrate leukocyte arrest in relation to the endothelium and basement membrane at 1.5 times the xy image. Scale bar = 50 μm. B : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in FVB/n mice). C : schematic of quantification of site of leukocyte blockade. Leukocytes were scored as being in 1 of 6 positions: 1 ) luminal, 2 ) apically arrested, 3 ) arrested partway through the endothelium, 4 ) arrested at the level of the basement membrane, 5 ) actively migrating through the basement membrane, or 6 ) fully extravasated. D : quantification of the site of leukocyte arrest in FVB/n mice. E : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in C57BL/6 mice). F : quantification of the site of leukocyte arrest in C57BL/6 mice. G : schematic model of anti-PECAM and anti-CD99 arrest in FVB/n and C57BL/6 mice. One hundred to 200 cells were analyzed per ear. Carrier-alone stimulation resulted in minimal leukocyte recruitment (data not shown). Total leukocytes per field of view, vessel length, and vessel diameter were equivalent for all treatment groups (data not shown). Data are representative of recordings from ≥4 mice per group with ≥8 images quantified per mouse. * P

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: 4D intravital microscopy uncovers critical strain differences for the roles of PECAM and CD99 in leukocyte diapedesis

    doi: 10.1152/ajpheart.00289.2016

    Figure Lengend Snippet: PECAM and CD99 function past the level of the endothelium in C57BL/6 mice. A : wild-type FVB/n and C57BL/6 mice were pretreated with control IgG, anti-PECAM, or anti-CD99 (3 mg/kg) prior to ear stimulation with croton oil (1% solution, 20 μl/ear) or carrier alone. After 4 h, ear tissue was collected and immunohistochemical staining was performed for PECAM [endothelial cells (ECs), red], myeloid-related protein 14 (neutrophils, green), and collagen IV [basement membrane (BM), purple]. 3-Dimensional confocal images were acquired from each sample and subsequently analyzed. Insets : z -orthogonal view along the indicated line to demonstrate leukocyte arrest in relation to the endothelium and basement membrane at 1.5 times the xy image. Scale bar = 50 μm. B : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in FVB/n mice). C : schematic of quantification of site of leukocyte blockade. Leukocytes were scored as being in 1 of 6 positions: 1 ) luminal, 2 ) apically arrested, 3 ) arrested partway through the endothelium, 4 ) arrested at the level of the basement membrane, 5 ) actively migrating through the basement membrane, or 6 ) fully extravasated. D : quantification of the site of leukocyte arrest in FVB/n mice. E : quantification of findings in A (percent leukocyte extravasation within 50 μm of postcapillary venules in C57BL/6 mice). F : quantification of the site of leukocyte arrest in C57BL/6 mice. G : schematic model of anti-PECAM and anti-CD99 arrest in FVB/n and C57BL/6 mice. One hundred to 200 cells were analyzed per ear. Carrier-alone stimulation resulted in minimal leukocyte recruitment (data not shown). Total leukocytes per field of view, vessel length, and vessel diameter were equivalent for all treatment groups (data not shown). Data are representative of recordings from ≥4 mice per group with ≥8 images quantified per mouse. * P

    Article Snippet: Rabbit anti-mouse collagen IV, rat anti-mouse myeloid-related protein 14 (MRP14), and rabbit anti-mouse α-smooth muscle actin (α-SMA) were purchased from Abcam (Cambridge, MA).

    Techniques: Mouse Assay, Immunohistochemistry, Staining

    Rab5 regulates cytochrome c release during H 2 O 2 -induced stress. ( A ) Live-cell imaging of a BAC GFP-Rab5 cell stained with 100 nM Mito-Red before (0 min) and after treatment (60 min) with 250 μM H 2 O 2 at 37°C. Inset images are shown and arrowheads indicate GFP-Rab5 recruitment to MOMP events (right panel). Scale bars, 10 μm. ( B ) HeLa cells over-expressing either GFP or GFP-Rab5 were treated with 250 μM H 2 O 2 at 37°C for 1 to 4 hr. Protein samples from cytosolic fractions were obtained and immunoblotted for cytochrome c and tubulin (as a loading control). ( C ) Densitometric quantification of cytochrome c release in ( B ). Data were collected from three independent experiments. Y-axis corresponds to the normalized ratio intensity of cytochrome c to the tubulin loading control. p-Values based on two-tailed t-tests. 10.7554/eLife.32282.034 Numerical data corresponding to the line traces presented in Figure 7C .

    Journal: eLife

    Article Title: Rab5 and Alsin regulate stress-activated cytoprotective signaling on mitochondria

    doi: 10.7554/eLife.32282

    Figure Lengend Snippet: Rab5 regulates cytochrome c release during H 2 O 2 -induced stress. ( A ) Live-cell imaging of a BAC GFP-Rab5 cell stained with 100 nM Mito-Red before (0 min) and after treatment (60 min) with 250 μM H 2 O 2 at 37°C. Inset images are shown and arrowheads indicate GFP-Rab5 recruitment to MOMP events (right panel). Scale bars, 10 μm. ( B ) HeLa cells over-expressing either GFP or GFP-Rab5 were treated with 250 μM H 2 O 2 at 37°C for 1 to 4 hr. Protein samples from cytosolic fractions were obtained and immunoblotted for cytochrome c and tubulin (as a loading control). ( C ) Densitometric quantification of cytochrome c release in ( B ). Data were collected from three independent experiments. Y-axis corresponds to the normalized ratio intensity of cytochrome c to the tubulin loading control. p-Values based on two-tailed t-tests. 10.7554/eLife.32282.034 Numerical data corresponding to the line traces presented in Figure 7C .

    Article Snippet: Cells were immunostained with the corresponding primary antibodies: anti-rabbit Rabenosyn-5/ZFYVE20 (Sigma Aldrich: HPA044878), anti-mouse EEA1 (BD Biosciences: 610457), anti-rabbit TOM20 (Santa Cruz Biotechnology: sc-11415), anti-rabbit APPL1 (Abcam: ab59592), anti-mouse APPL2 (home-made), anti-mouse Rab5 (BD Biosciences: 610724), anti-mouse cytochrome c (Abcam: ab6311), and anti-rabbit Alsin (Novus Biological: NBP2-14284) antibodies.

    Techniques: Live Cell Imaging, BAC Assay, Staining, Expressing, Two Tailed Test

    Schematic model depicting the role of Rab5-Alsin-mitochondria during oxidative stress. In the normal condition, mitochondria (Mito, red) are elongated and tubular (left). Rab5 (green) are localized on early endosomes (EE) to assemble the Rab5 machinery for endosomal maturation and membrane trafficking. At steady state, some EE make transient contacts with mitochondria. During oxidative stress (e.g. laser- or chemically-induced), mitochondria undergo MOMP and a dramatic morphological transformation into rounded and swollen structures (right). The release of the apoptotic factor, cytochrome c, from mitochondria into the cytosol is associated with a re-localization event of Rab5 from EE to mitochondria via a cytosolic intermediate, accompanied by an increase in EE-mitochondria MCS. The recruitment and activation of Rab5 on mitochondria depend on the Rab5 GEF Alsin (blue), which leads to a selective recruitment of Rabenosyn-5 (light green). This signaling cascade on mitochondria is a reversible process that regulates the apoptotic program (e.g. cytochrome c release and caspase activation) and thus, promotes overall cell survival.

    Journal: eLife

    Article Title: Rab5 and Alsin regulate stress-activated cytoprotective signaling on mitochondria

    doi: 10.7554/eLife.32282

    Figure Lengend Snippet: Schematic model depicting the role of Rab5-Alsin-mitochondria during oxidative stress. In the normal condition, mitochondria (Mito, red) are elongated and tubular (left). Rab5 (green) are localized on early endosomes (EE) to assemble the Rab5 machinery for endosomal maturation and membrane trafficking. At steady state, some EE make transient contacts with mitochondria. During oxidative stress (e.g. laser- or chemically-induced), mitochondria undergo MOMP and a dramatic morphological transformation into rounded and swollen structures (right). The release of the apoptotic factor, cytochrome c, from mitochondria into the cytosol is associated with a re-localization event of Rab5 from EE to mitochondria via a cytosolic intermediate, accompanied by an increase in EE-mitochondria MCS. The recruitment and activation of Rab5 on mitochondria depend on the Rab5 GEF Alsin (blue), which leads to a selective recruitment of Rabenosyn-5 (light green). This signaling cascade on mitochondria is a reversible process that regulates the apoptotic program (e.g. cytochrome c release and caspase activation) and thus, promotes overall cell survival.

    Article Snippet: Cells were immunostained with the corresponding primary antibodies: anti-rabbit Rabenosyn-5/ZFYVE20 (Sigma Aldrich: HPA044878), anti-mouse EEA1 (BD Biosciences: 610457), anti-rabbit TOM20 (Santa Cruz Biotechnology: sc-11415), anti-rabbit APPL1 (Abcam: ab59592), anti-mouse APPL2 (home-made), anti-mouse Rab5 (BD Biosciences: 610724), anti-mouse cytochrome c (Abcam: ab6311), and anti-rabbit Alsin (Novus Biological: NBP2-14284) antibodies.

    Techniques: Transformation Assay, Activation Assay