Ab51891

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab51891 is an antibody product offered by Abcam. It is a recombinant monoclonal antibody that can be used in various laboratory applications. The core function of this product is to bind to and detect a specific target molecule.

Automatically generated - may contain errors

Lab products found in correlation

Lsm 510, by Zeiss (1 mentions) Caspase 3, by Cell Signaling Technology (1 mentions) Ab53226, by Abcam (1 mentions) Sc 13979, by Santa Cruz Biotechnology (1 mentions) Mouse anti vimentin cy3, by Merck Group (1 mentions) Dmi4000d microscope, by Leica (1 mentions) Iblot dry blotting system, by Thermo Fisher Scientific (1 mentions) Sc 1616 r, by Santa Cruz Biotechnology (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Chemidoc xrs gel imaging system, by Bio-Rad (1 mentions) Bs3489, by Bioworld Technology (1 mentions) Ab150165, by Abcam (1 mentions) Ap0707, by ABclonal (1 mentions) Bca protein quantitative detection kit, by Beyotime (1 mentions) Sa00001 2, by Proteintech (1 mentions) Ab150077, by Abcam (1 mentions)

5 protocols using ab51891

Immunofluorescence analysis of lung cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary AEC isolated from lung biopsy were fixed in 3% paraformaldehyde (PFA) for 10 minutes at room temperature, washed three times in 0.1% glycin/PBS, and incubated with either rabbit polyclonal antibody to E-cadherin (1:200) (#ab53226) (Abcam, USA), or rabbit polyclonal SP-C (1:50) (#sc13979) (Santa Cruz Biotechnology, USA) or mouse monoclonal antibody to surfactant protein A (SP-A) (1:200) (#ab51891) (Abcam, USA) over night in a moist chamber at 4 °C. For detection Cy3 labelled anti rabbit secondary antibody was used for E-cadherin and SP-C, while a alkaline phosphatase labeling was used to detect SP-A. A549 cells were treated with TN or TG, for 18 hours and 7 days respectively and grown in either BMSC-cm or normal media for 24 hours. Cells were then fixed in 3% PFA as described above and subsequently stained overnight at 4 °C in a moist box with anti-cytokeratin, pan-FITC anti-mouse (1:200) (#C5992), mouse anti-vimentin-Cy3 (1:1000) (both: Sigma, USA) (#C9080) or caspase-3 (1:200) (#9662 S) (Cell Signaling, USA). Samples were then washed 3 times with PBS and analyzed under a confocal microscope LSM 510 Carl Zeiss or under Leica DMI4000D microscope.

Nita I., Hostettler K., Tamo L., Medová M., Bombaci G., Zhong J., Allam R., Zimmer Y., Roth M., Geiser T, & Gazdhar A. (2017). Hepatocyte growth factor secreted by bone marrow stem cell reduce ER stress and improves repair in alveolar epithelial II cells. Scientific Reports, 7, 41901.

+ Open protocol

+ Expand

Quantification of Surfactant Proteins A and D

Check if the same lab product or an alternative is used in the 5 most similar protocols

After a post-exposure recovery period, ATII cells were lysed using Cell Lytic M (with protease inhibitor cocktail), incubated on ice for 30min and then centrifuged at 3000g for 20min at 4°C. Protein concentrations from ATII extracts were determined using Bradford Reagent. Equal volume of samples (mixed with 4X NuPAGE LDS sample loading buffer and 10X NuPAGE sample reducing agent, then boiled at 100°C) were loaded into NuPage 4–12% gels and electrophoretically separated using sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Following gel electrophoresis, gels were rinsed in distilled water then transferred to the nitrocellulose membrane using the i-Blot^™ Dry Blotting System (Invitrogen, UK). SP-A and SP-D protein on this membrane was then detected using the anti-human SP-A antibody, ab51891, and the anti-human SP-D antibody, ab97849 (both from Abcam, Cambridge, UK). SPA and SP-D protein bands were imaged using Ultraviolet Transilluminator GelDoc-It and densitometric quantification was performed using Visionworks.

Sweeney S., Theodorou I.G., Zambianchi M., Chen S., Gow A., Schwander S., Zhang J.(., Chung K.F., Shaffer M.S., Ryan M.P., Porter A.E, & Tetley T.D. (2015). Silver nanowire interactions with primary human alveolar type-II epithelial cell secretions: contrasting bioreactivity with human alveolar type-I and type-II epithelial cells. Nanoscale, 7(23), 10398-10409.

+ Open protocol

+ Expand

Immunochemical Analysis of Alveolar SP-A and TTF-1

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunochemistry was performed by 2 investigators in a blinded manner. SP-A expression in situ and distribution were assessed in different lung cells, including PNII and alveolar macrophages (MACR). Thyroid transcription factor-1 (TTF-1) is mainly found in the thyroid, lung, and brain tissues in humans,^{[17 (link),18 (link)]} and mostly located in nuclei of PNII in the lung. A study has tagged TTF-1 to count type II alveolar epithelial cells.^{[19 (link)]} SP-A and TTF-1 in formalin-fixed, paraffin-embedded lung tissue sections were detected by the SP method. Primary mouse anti-human-TTF-1 (Santa Cruz, sc-53136, 1:200 dilution) and mouse anti-human-SP-A (Abcam, ab51891, 1:1000 dilution) antibodies were used to assess total PNII and SP-A+ PNII, respectively. Bovine serum albumin was used as negative control. Sections were counterstained with Mayer hematoxylin. SP-A optical density (OD) per square millimeter semiquantitatively reflected total SP-A expression in situ.^{[20 (link)]} The rates of SP-A+ PNII and MACR reflected SP-A distribution in the lung tissue. SP-A expression was determined in 20 randomly selected high power fields.

Liu Z., Chen S., Xu Y., Liu X., Xiong P, & Fu Y. (2020). Surfactant protein A expression and distribution in human lung samples from smokers with or without chronic obstructive pulmonary disease in China. Medicine, 99(7), e19118.

+ Open protocol

+ Expand

Quantifying Lung SP-A Protein Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total SP-A protein expression levels in the lung tissue were assessed, with β-actin as an internal reference. Lung tissue lysates were obtained and equal amounts total protein (50 μg) were separated by 12.5% Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophorosis under reducing conditions. The protein bands were then transferred onto Polyvinylidene fluoride membranes. Detection was performed with mouse anti-human-SP-A (Abcam, ab51891, 1:2000 dilution) and rabbit anti-human-actin (Santa Cruz, sc-1616-R; 1:1500 dilution) antibodies. After film exposure, the bands were quantified with the Image Pro Plus 6.0 software, based on integrated optical density (IOD) values.

+ Open protocol

+ Expand

Western Blot Analysis of Immune Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tissue samples were lysed using RIPA lysis buffer (P0013B, Beyotime, Beijing, China), followed by 140000×g centrifugation at 4°C for 15 min on the ice. Then, supernatant was collected. The concentration of extracted protein was determined using BCA protein quantitative detection kit (P0009, Beyotime). Protein samples were subjected on to SDS/PAGE and transferred to PVDF membrane. After that, membrane was blocked using 5% milk/TBST at room temperature for 1 h. Membrane was incubated with 30 μg primary antibodies against SP-A (1:1000; ab51891, Abcam, Cambridge, U.K.), IKBα (1:1500; 10268-1-AP, Proteintech, Chicago, U.S.A.), p-IKBα (1:1000; AP0707, ABclonal, Wuhan, China), TLR4 (1:600; BS3489, Bioworld, Minnesota, U.S.A.) and GAPDH (1:5000, SA00001-2, Proteintech, Wuhan, China) at 4°C overnight, followed by incubation with horseradish peroxidase-labeled goat anti-rabbit secondary antibody (1:5000, ab150077, Abcam) and goat anti-rat secondary antibody (1:5000, ab150165, Abcam) at room temperature for 1 h. Protein blots were visualized using Enhanced Luminol reagent and oxidizing reagent. The results were observed using ChemiDoc™ XRS+ Gel imaging system (Bio-Rad, Shanghai, China).

Qian J., Li G., Jin X., Ma C., Cai W., Jiang N, & Zheng J. (2020). Emodin protects against intestinal and lung injury induced by acute intestinal injury by modulating SP-A and TLR4/NF-κB pathway. Bioscience Reports, 40(9), BSR20201605.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!