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Anti cd71

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Anti-CD71 is a monoclonal antibody that recognizes the transferrin receptor (CD71), a cell surface protein involved in cellular iron uptake. It is commonly used as a marker for cell proliferation and activation.

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Lab products found in correlation

Glutamax, by Thermo Fisher Scientific (3 mentions) Tcs sp8 aobs confocal microscope, by Leica (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Anti cd8 apc cy7, by BD (1 mentions) Anti cd56 pe cf594, by BD (1 mentions) Anti rab5, by Cell Signaling Technology (1 mentions) Anti rab8, by Cell Signaling Technology (1 mentions) Anti rab27a, by Abcam (1 mentions) Anti syntaxin6, by Cell Signaling Technology (1 mentions) Anti vamp3, by Abcam (1 mentions) Anti snap23, by Abcam (1 mentions) Goat anti rabbit af647, by Thermo Fisher Scientific (1 mentions) Chicken anti goat af647, by Thermo Fisher Scientific (1 mentions) Anti rab7, by Santa Cruz Biotechnology (1 mentions) Anti granzyme b, by BD (1 mentions) Anti rab11a, by Cell Signaling Technology (1 mentions) Anti rab37, by Abcam (1 mentions) Nitrocellulose filter membrane, by Merck Group (1 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Anti xct, by Cell Signaling Technology (1 mentions)

4 protocols using anti cd71

1

Transferrin Receptor Recycling in Fibroblasts

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The transferrin receptor recycling protocol was adapted from the literature [49 (link), 76 (link)]. Fibroblasts were plated on glass coverslips 48 h before the experiment. After rinsing with PBS, fibroblasts were kept at 4 °C for 30 min in serum-free DMEM/GlutaMax (Gibco, Thermofisher Scientific). Fibroblasts were then rinsed on ice and incubated in DMEM / GlutaMax / SVF 10% / penicillin-streptomycin 1% supplemented with transferrin (100 μg / mL, T13342, Thermofisher Scientific) at 37 °C for 15 min. Cells were rinsed on ice and fixed in PFA 4% for 10 min. Cells were then immunolabelled against the surface transferrin receptor, therefore with an immunolabelling protocol without permeabilization. Apart from permeabilization, the staining protocol is identical to that presented above. Cells incubated 1 h in primary antibody anti-CD71 (monoclonal rabbit, 13,208, Cell Signaling, 1/300) and 1 h in secondary antibody anti-mouse Alexa Fluor 647 (donkey, A− 31,571, ThermoFisher Scientific, 1/1000) were then observed under a Leica TCS SP8 AOBS confocal microscope with an oil immersion objective 63x / NA = 1.40 on the ICMQuant platform.
Botté A., Lainé J., Xicota L., Heiligenstein X., Fontaine G., Kasri A., Rivals I., Goh P., Faklaris O., Cossec J.C., Morel E., Rebillat A.S., Nizetic D., Raposo G, & Potier M.C. (2020). Ultrastructural and dynamic studies of the endosomal compartment in Down syndrome. Acta Neuropathologica Communications, 8, 89.
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2

Imaging Flow Cytometry for Immune Cell Analysis

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For imaging flow cytometry experiments: anti-CD8 APC-Cy7 (BD Biosciences, San Jose, CA or Tonbo Biosciences, San Diego, CA) and anti-CD56 PE-CF594 (BD Biosciences) were used for surface staining; all intracellular stains included anti-perforin D48 PE or BV421 (Biolegend, San Diego, CA) and anti-perforin δG9 FITC (Biolegend) and one of the following unconjugated antibodies: anti-CD71 (Cell Signaling Technology, Danvers, MA), anti-granzyme B (BD Biosciences), anti-rab3D (Abcam, Cambridge, MA), anti-rab4 (Pierce Antibodies, Waltham, MA), anti-rab5 (Cell Signaling Technology), anti-rab7 (Santa Cruz Biotechnology, Dallas, TX), anti-rab8 (Cell Signaling Technology), anti-rab11a (Cell Signaling Technology), anti-rab27a (Abcam), anti-rab35 (Abcam), anti-rab37 (Abcam), anti-syntaxin6 (Cell Signaling Technology), anti-syntaxin7 (R&D systems, Minneapolis, MN), anti-vti1b (Abcam), anti-VAMP3 (Abcam), anti-VAMP4 (Abcam), or anti-SNAP23 (Abcam). The unconjugated antibodies were detected using goat anti-rabbit AF647, chicken anti-goat AF647, or donkey anti-sheep AF647 secondary antibodies (Invitrogen, Carlsbad, CA). For confocal microscopy, perforin D48 FITC (Abcam), perforin δG9 AF647 (Biolegend), and goat anti-rabbit or donkey anti-sheep AF568 secondary antibodies (Invitrogen) were used, along with the selected rab or SNARE antibody.
Lesteberg K.E., Orange J.S, & Makedonas G. (2017). Recycling endosomes in human cytotoxic T lymphocytes constitute an auxiliary intracellular trafficking pathway for newly synthesized perforin. Immunologic research, 65(5), 1031-1045.
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3

Protein Expression Analysis by Western Blot

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RIPA buffer containing a protease inhibitor (Beyotime, Nantong, China) was used to lyse the tissues or cells. Equal amounts of protein were separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (NCM Biotech, Suzhou, China) and then transferred onto nitrocellulose filter membranes (Millipore, USA). The membranes were blocked with 5% skim milk for 2 h. After washing twice with Tris-Buffered Saline Tween-20 buffer (Sevicebio, Wuhan, China), the membranes were incubated with primary antibodies, including anti-Fatty Acid Synthase (1:1000; Cell Signaling Technology), anti-GAPDH (1:5000; Proteintech), anti-xCT (1:1000; Cell Signaling Technology), anti-GPX4 (1:1000; Cell Signaling Technology), anti-CD71 (1:1000; Cell Signaling Technology), anti-p-ERK (1:1000; Cell Signaling Technology), or anti-ERK (1:1000; Cell Signaling Technology) overnight, followed by anti-rabbit IgG (1:2000; Cell Signaling Technology) or anti-mouse IgG (1:5000; Abmart) for 2 h. Lastly, the images were developed with ChemiScope 6100 (Clinx, Shanghai, China) after adding chemiluminescent substrate (NCM biotech).
Ye M., Lu F., Chen J., Yu P., Xu Y., He N., Hu C., Zhong Y., Yan L., Gu D., Xu L., Bai J., Tian Y, & Tang Q. (2023). Orlistat Induces Ferroptosis in Pancreatic Neuroendocrine Tumors by Inactivating the MAPK Pathway. Journal of Cancer, 14(8), 1458-1469.
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4

Comprehensive Cell Signaling Antibody Panel

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Cell Signaling: anti-CD71 (#13113), anti-cleaved Caspase 3 (#9661S), anti-His (#2365), anti-IκBα (#4814), anti-Integrinα6 (#3750), anti-STX6 (#2869), anti-TNF-R1 (#3736), anti-PARP (#9542S).
ENZO: anti-Ceramide (15B4; ALX-804-196-T050).
LSbio: anti-Lypla2/APT2 (LS-C158086).
LifeTechnologies: anti-mouse Alexafluor488 (A21202).
Millipore: anti-mouse light chain (AP200P), anti-rabbit light chain (MAB201P).
Proteintech: anti-βActin (HRP-60008), anti-GAPDH (HRP-60004).
Santa Cruz Biotechnology: anti-Rab5B (sc-598).
Sigma-Aldrich: anti-PPT1 (HPA021546).
ThermoFisher: anti-Lypla1/APT1 (PA5–28034).
Zingler P., Särchen V., Glatter T., Caning L., Saggau C., Kathayat R.S., Dickinson B.C., Adam D., Schneider-Brachert W., Schütze S, & Fritsch J. (2019). Palmitoylation is required for TNF-R1 signaling. Cell Communication and Signaling : CCS, 17, 90.
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