Whole cell lysates were prepared and loaded onto SDS-PAGE. After transfer, the membrane was blocked with 5% skim milk for 1hr and then incubated in diluted primary antibodiesATP11B (ab05377), ATP7B (ab124973), tubulin (ab210797) or GAPDH (ab181602), obtained from Abcam, Cambridge, USA) overnight. The membrane was washed with TBS-T three times and incubated with an HRP-labeled secondary antibody for 1hr. The membrane was developed using a Bio-Rad developer system.
Ab210797
Manufactured by Abcam
Sourced in United States
Ab210797 is a primary antibody that targets the protein of interest. It is suitable for use in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Developer system, by Bio-Rad (2 mentions)
Ab194486, by Abcam (2 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (2 mentions)
Ab181602, by Abcam (1 mentions)
Ab124973, by Abcam (1 mentions)
Ab178945, by Santa Cruz Biotechnology (1 mentions)
Ab32575, by Abcam (1 mentions)
Ab222783, by Abcam (1 mentions)
Ab76067, by Abcam (1 mentions)
Ab199956, by Abcam (1 mentions)
Ab8245, by Abcam (1 mentions)
Ab1316, by Abcam (1 mentions)
Gapdh, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Ecl detection kit, by Santa Cruz Biotechnology (1 mentions)
Mammalian cell lysis reagent, by Thermo Fisher Scientific (1 mentions)
Ab17147, by Abcam (1 mentions)
Ab92381, by Abcam (1 mentions)
Ab270264, by Abcam (1 mentions)
Ab231036, by Abcam (1 mentions)
10 protocols using ab210797
1
Western Blot Protein Analysis
2
Quantification of Vascular and Nitric Oxide Proteins
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Western blot (WB) was performed as previously described.21 Primary antibodies include anti‐CD31 (Abcam, ab222783, 1:2000), anti‐VEGFA (Abcam, ab1316, 5 µg/mL), anti‐neuronal nitric oxide synthase (nNOS, Abcam, ab76067, 1:1000), anti‐endothelial nitric oxide synthase (eNOS, Abcam, ab199956, 1:1000), anti‐inducible nitric oxide synthase (iNOS, Abcam, ab178945, 1:1000), anti‐SOD (Santa Cruz Biotechnology, sc‐101523, 1:500), anti‐α‐SMA (Abcam, ab32575, 1:2500). Anti‐GAPDH (Abcam, ab8245, 1:5000) and anti‐tubulin (Abcam, ab210797, 1:1000) were used as the internal standard. The image of protein expression was detected by Odyssey CLX Two‐colour infrared laser imaging system (LI‐COR Biosciences). Densitometric analysis of the bands was implemented using ImageJ software (National Institutes of Health).
3
Western Blot Protein Detection
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Whole cell lysates were prepared and loaded onto sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After transfer, the membrane was blocked with 5% skim milk in Tris-buffered saline (TBS) supplemented with 0.05% Tween 20 (TBST) for 1 hour and then incubated in diluted primary antibodies PKR, p90RSK, tubulin (#ab210797) or GAPDH (#ab181602) (obtained from Abcam, Cambridge, USA) overnight. The membrane was washed with TBS-T three times and incubated with an horseradish peroxidase (HRP)-labeled secondary antibody for 1 hour. The membrane was developed using a Bio-Rad developer system.
4
Immunofluorescence Staining of Lung Tissue
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IF staining was performed as previously reported 20 (link). Paraffin sections of the lung tissue and cell slides were incubated with α-SMA (1184-1, Eptomics, Burlingame, CA, USA, 1:300), Ac-α-Tub (sc-23950, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1:200), γ-Tubulin (ab210797, Abcam, Cambridge, MA, USA, 1:200) and ADP-ribosylation factor-like protein 13B (ARL13B) (GTX122703, GeneTex, Inc., Irvine, CA, USA, 1:200). The slides were then counterstained with DAPI (5 g/L, Beyotime, Haimen, China) and were visualised using an Olympus DP80 microscope (Olympus Soft Imaging).
5
Protein Extraction and Western Blot Analysis
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We applied the Mammalian Cell Lysis Reagent (Thermo Scientific, Rockford, IL) to extract proteins from cells and tissues according to the protocol. The protein was separated by 10% SDS‐PAGE and then transferred to PVDF membranes. Then blocked with 5% non‐fat milk and incubated with the primary antibodies and followed by the incubation with HRP‐conjugated secondary antibodies. Then detect the protein bands using ECL detection kit (Santa Cruz, CA). Primary antibodies of anti‐ERK1/2 (9102), p‐ERK1/2 (4370), NF‐κB (p65) (8242), p‐NF‐κB (p65) (3033), c‐Fos (4384), FosB (2263), c‐Jun (9165), CDK6 (3136), Cyclin D1 (2922), Cyclin D3 (2936) and P27 (3686) were purchased from Cell Signaling Technology (Boston, MA); anti‐FRA1 (ab124722), CDK4 (ab108357), P15 (ab53034), mindin (ab187920), GAPDH (ab181602) and tubulin (ab210797) from were purchased Abcam (Cambridge, MA), and HRP‐conjugated anti‐rabbit (A8275) and mouse (A5906) secondary antibodies were purchased from Sigma‐Aldrich (St. Louis, MO).
6
Comprehensive Immunophenotyping Assay
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The following primary antibodies were used: anti-tubulin (Abcam, ab210797), anti-KMT5A (Abcam, ab111691), anti-CD8 (Abcam, ab17147), anti-CD8 (Abcam, ab217344), anti-CD3 (Abcam, ab16669), anti-ubiquitin (CST, #43124), anti-CD69 (Abcam, ab54217), anti-FLAG (Abcam, ab205606), anti-HA (Abcam, ab236632), anti-6X His (Abcam, ab213204), anti-CD73 (Abcam, ab54217), anti-CD73 (Abcam, ab54217), anti-COP1 (Abcam, ab56400), anti-MKRN1 (Abcam, ab72054), anti-MDM2 (BOSTER, BA3612–2), and anti-Ki-67 (CST, #12202), anti-CD69 (BOSTER, A00529–2), anti-IFN-γ (Abcam, ab231036), anti-IL-2 (Abcam, ab92381), anti-TNF-α (Abcam, ab270264), anti-perforin (Abcam, ab47225), anti-Granzyme B (BOSTER, A00353), anti-perforin (Abcam, ab16074). The following secondary antibodies were used: goat anti-mouse (CST) and goat anti-rabbit (CST). MG132 and cycloheximide (CHX) were purchased from CST (USA). The CMV peptide pool stimulating CD8+ T cells was obtained from Mabtech (Sweden).
7
Comprehensive Protein Expression Analysis
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Anti-CAV3 (Abcam, ab289544, rabbit recombinant multiclonal, 1:1000), anti-NDUFA10 (Abcam, ab174829, rabbit monoclonal, 1:1000), anti-BAX (Abmart, M20008S, rabbit monoclonal, 1:1000), anti-Bcl2 (Abmart, T40056S, rabbit monoclonal, 1:1000), anti-cleaved caspase-3 (Cell Signalling, 9669 s, rabbit monoclonal, 1:1000), anti-SOD2 (ABclonal, A1340, rabbit monoclonal, 1:1000), anti-Cytochrome c (proteintech, 66264-1-Ig, mouse monoclonal, 1:5000), anti-VDAC1/Porin (proteintech, 55259-1-AP, rabbit polyclonal, 1:1000), anti-ATP1A1 (proteintech, 14418-1-AP, rabbit polyclonal, 1:5000), anti-Lamin B1 (proteintech, 12987-1-AP, rabbit polyclonal, 1:5000), anti-PDI (proteintech, 11245-1-AP, rabbit polyclonal, 1:500), anti-β-actin (ABclonal, AC026, rabbit monoclonal, 1:1000), anti-α-tubulin (Abcam, ab210797, mouse polyclonal, 1:1000), HRP-conjugated anti-rabbit or anti-mouse IgG (ABclonal, AS014, AS003, polyclonal, 1:5000), and HRP-conjugated anti-rabbit IgG- heavy chain or light chain (ABclonal, AS063, AS061, polyclonal, 1:5000) antibodies were used for WB analysis.
8
Quantifying Endothelial Protein Expression
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To determine the protein expression levels of occludin and KLF2, HBMVECs were lysed using radio-immunoprecipitation assay (RIPA) buffer supplemented with protease inhibitor cocktail after undergoing the indicated treatment. The proteins were then separated by size by loading 20 μg total cell lysates onto 4–20% precasted polyacrylamide gel electrophoresis (PAGE) gels, and then transferring the separated protein mixture onto polyvinylidene fluoride (PVDF) membranes. Specific antibodies were used to determine the protein expression levels. The following antibodies were used in this study: p-ERK5 (1 : 500, #3371, Cell Signaling Technology, USA); ERK5 (1 : 1000, #12950, Cell Signaling Technology, USA); KLF2 (1 : 1000, #ab194486, Abcam, USA); occludin (1 : 500, #ab31721, Abcam, USA); tubulin (1 : 10 000, #ab210797, Abcam, USA); anti-rabbit IgG, HRP-linked antibody (1 : 2000, #7074, Cell Signaling Technology, USA); anti-mouse IgG, HRP-linked Antibody (1 : 2000, #7076, Cell Signaling Technology, USA).
9
Western Blot Analysis of Protein Expression
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After the indicated treatment, HAECs were lysed using RIPA buffer supplemented with protease inhibitor cocktail. Then, 20 μg cell lysates were loaded onto 4–20% precasted PAGE gels to separate the proteins by size. The separated protein mixture was then transferred onto PVDF membranes and the corresponding protein levels were detected using their specific antibodies. Blots were quantified by densitometry analysis of three exposures using TotalLab TL100 image analysis software. The following antibodies were used in this study: p-ERK5 (1:500, #3371, Cell signaling technology, USA); ERK5 (1:2000, #ab196609, Abcam, USA); KLF2 (1:1000, #ab194486, Abcam, USA); tubulin (1:10,000, #ab210797, Abcam, USA); anti-rabbit IgG, HRP-linked Antibody (1:2000, #7074, Cell Signaling Technology, USA); anti-mouse IgG, HRP-linked Antibody (1:2000, #7076, Cell Signaling Technology, USA).
10
Protein Extraction and Western Blot Analysis
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Total proteins from human tissue or glioma cells were extracted by using a radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime Biotechnology Co., Ltd., Shanghai, China). The protein samples were separated using 8% SDS-PAGE. After transferred onto PVDF membrane, the primary antibodies were added: anti-LRIG1 (PA5-52860, 1:500, Invitrogen, Carlsbad, CA, USA), anti-Akt (ab235958, 1:1000, Abcam, UK), anti-Akt (phospho S473) (ab81283, 1:1000, Abcam, UK), anti-CTLA-4(PA5-47547, 1:250, Invitrogen, Carlsbad, CA, USA), anti-Tubulin (ab210797, 1:1000, Abcam, UK). Tubulin served as internal control. After 24 hours of incubation, membrane was incubated along with secondary antibody immunoglobulin G (IgG) at room temperature for 2 hours.
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