Anti cd99

Manufactured by Abcam

Sourced in United Kingdom

Anti-CD99 is a primary antibody that targets the CD99 cell surface antigen. CD99 is a transmembrane glycoprotein expressed on various cell types. This antibody can be used for the detection and analysis of CD99-expressing cells in research applications.

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Lab products found in correlation

Anti erk1 phospho t202 erk2 phospho t185, by Abcam (2 mentions) Anti cyclin a2, by Abcam (1 mentions) Anti cyclin e1, by Abcam (1 mentions) Anti p21, by Abcam (1 mentions) Anti cdk2, by Abcam (1 mentions) Anti erk1 erk2, by Abcam (1 mentions) Anti hif 1α, by Cell Signaling Technology (1 mentions) Triton x 100, by Sangon (1 mentions) Alexa fluor 647, by Abcam (1 mentions) Lsm 710 confocal laser scanning microscope, by Zeiss (1 mentions) Supersignal west pico substrate, by Thermo Fisher Scientific (1 mentions) Anti rela p65, by Santa Cruz Biotechnology (1 mentions) Anti actin, by Merck Group (1 mentions) Iblot transfer system, by Thermo Fisher Scientific (1 mentions) Anti calnexin, by LifeSpan BioSciences (1 mentions) Anti pan akt clone c67e7, by Cell Signaling Technology (1 mentions) Anti irf3 clone fl 425, by Santa Cruz Biotechnology (1 mentions) Bis tris polyacrylamide gel, by Thermo Fisher Scientific (1 mentions) Anti fat1, by Merck Group (1 mentions) Image studio software, by LI COR (1 mentions)

4 protocols using anti cd99

Western Blotting Procedure for Protein Analysis

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Western blotting was conducted as previously described [17 (link)] using anti-HIF-1α (1:1000; Cell Signaling Technology, Danvers, MA, United States), anti-cyclin E1 (1:1000; Abcam, Cambridge, United Kingdom), anti-cyclin A2 (1:1000; Abcam), anti-CDK2 (1:1000; Abcam), anti-p21 (1:1000; Abcam), anti-CD99 (1:1000; Abcam), anti-ERK1+ERK2 (1:5000; Abcam), anti-ERK1 (phospho T202)+ERK2 (phospho T185) (1:1000; Abcam) and anti-GAPDH (1:1000; Sangon Biotech Corp., Shanghai, China) primary antibodies. The membranes were then incubated with secondary anti-rabbit antibodies conjugated to horseradish peroxidase (1:4,000; Abcam). Proteins were detected using Pierce^TMenhanced chemiluminescence (ECL) western blot analysis substrate (Thermo Fisher Scientific Inc.) and analyzed with ImageJ software (NIH, Bethesda, MD, United States).

Feng X.D., Zhu J.Q., Zhou J.H., Lin F.Y., Feng B., Shi X.W., Pan Q.L., Yu J., Li L.J, & Cao H.C. (2021). Hypoxia-inducible factor-1α–mediated upregulation of CD99 promotes the proliferation of placental mesenchymal stem cells by regulating ERK1/2. World Journal of Stem Cells, 13(4), 317-330.

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Immunofluorescence Staining of CD99 and ERK

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Cells were grown on plates to about 70% confluence. After washing with PBS, the cells were fixed in 4% paraformaldehyde for 30 min in darkness and permeabilized with 0.3% Triton X-100 (Sangon Biotech Corp.) in PBS. Next, cells were blocked with 5% BSA and incubated with anti-CD99 (1:100; Abcam) and anti-ERK1 (phospho T202) +ERK2 (phospho T185) (1:100; Abcam) antibodies at 4 °C overnight. Goat anti-rabbit immunoglobulin G (Alexa Fluor^® 647, 1:200; Abcam) was incubated with the fixed cells at room temperature for 1 h and protected from light. 4′,6-Diamidino-2-phenylindole was used to counterstain nuclei. Stained slides were observed with a Zeiss LSM710 confocal laser-scanning microscope (Carl Zeiss AG, Germany).

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Western Blot Analysis of Immune Proteins

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Guanidine whole-cell lysates were precipitated using a ProteoExtract protein precipitation kit (Calbiochem). Proteins were re-dissolved in 2% SDS/Tris 200 mM pH8.5 with sonication. Protein concentration was measured by BCA (Pierce) then lysates were reduced with 100 mM DTT in loading buffer for 20 min at 65°C. Equal protein amounts were separated by SDS/PAGE using 4%–12% Bis-Tris polyacrylamide gels (Life Technologies), then transferred to PVDF membranes using the iBlot transfer system (Life Technologies). The following primary antibodies were used: anti-IFIT1 (clone 3G8, Lifespan), anti-IFIT3 (cat no. GTX112442, GeneTex), anti-MX1 (clone 2G12, GeneTex), anti-Calnexin (cat no. C142768, Lifespan), anti-MHC class I (clone HC-10, Abcam), anti-CD99 (cat no. ab108297, Abcam), anti-CLEC1A (cat no. AF1704, R and D systems), anti-FAT1 (cat no. HPA023882, Sigma), anti-PCDHGC3 (cat no. sc-134416, Santa Cruz), anti-IRF3 (clone FL-425, Santa Cruz), anti-NF-kB p50/105 (cat no. sc-7178, Santa Cruz), anti-p65/RELA (cat no. sc-109, Santa Cruz), anti-Pan-Akt (clone C67E7, Cell Signaling), anti-actin (cat no. A2066, Sigma). HRP-conjugated secondary antibodies were anti-rabbit (Cell Signaling), anti-mouse (Santa Cruz) or anti-goat (Santa Cruz). Reactive bands were detected by SuperSignal West Pico substrate (Thermo).

Weekes M.P., Tomasec P., Huttlin E.L., Fielding C.A., Nusinow D., Stanton R.J., Wang E.C., Aicheler R., Murrell I., Wilkinson G.W., Lehner P.J, & Gygi S.P. (2014). Quantitative Temporal Viromics: An Approach to Investigate Host-Pathogen Interaction. Cell, 157(6), 1460-1472.

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Immunoblotting Analysis of Cell Signaling Proteins

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Preparation of cellular lysates and subsequent immunoblotting were performed as previously described.^{54 (link)} The antibodies used for the immunoblotting are anti-CD99 (Abcam, # ab75858, EPR3097Y), anti-PARP (Cell Signaling Technology, #9542), anti-Caspase 3 (Cell Signaling Technology (CST), #9662), anti-H2A.X (CST, #7631, D17A3), anti-γH2A.X (CST, #9718, Ser139 - 20E3), anti-ACTIN-HRP (Abcam, #ab49900, AC-15), anti-Rabbit IgG-HRP (Cytiva, # NA934), and anti-Mouse IgG-HRP (Cytiva, # NA9311). Enhanced chemiluminescence (ECL) signal on PVDF membranes was captured by using LI-COR Odyssey XF imaging system (LI-COR Biosciences) and densitometry analysis of the protein bands were performed by using Image Studio software (LI-COR Biosciences).

Balaraman K., Deniz E., Nelson E., Pilicer S.L., Atasoy S., Molotkova A., Sevim H., Tiwari P.B., Üren A, & Wolf C. (2023). Design, Synthesis and Biological Evaluation of Nucleosidic CD99 Inhibitors that Selectively Reduce Ewing Sarcoma Viability. European journal of medicinal chemistry, 251, 115244.

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