Pac 1 antibody

Manufactured by BD

Sourced in United States

PAC-1 antibody is a laboratory research tool used for the detection and analysis of the PAC-1 protein. It is commonly used in various cell-based assays and immunohistochemical applications to study the expression and localization of PAC-1. The PAC-1 antibody is a specific and sensitive reagent for researchers working in the field of cell biology and molecular biology.

Automatically generated - may contain errors

Lab products found in correlation

β actin antibody, by Cell Signaling Technology (3 mentions) Fitc conjugated mouse anti human cd41a, by BD (3 mentions) Fitc conjugated goat anti mouse igg, by ZSGB-BIO (3 mentions) Pe conjugated anti human mouse cd62p p selectin, by Thermo Fisher Scientific (2 mentions) Anti human glycoprotein 6 purified antibody, by Thermo Fisher Scientific (2 mentions) Fitc conjugated anti cd42b antibody, by Abcam (2 mentions) Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (2 mentions) Thrombin, by Chrono-Log (2 mentions) Ab69507, by Abcam (1 mentions) Ez link nhs biotin, by Thermo Fisher Scientific (1 mentions) 9eg7 antibody, by BD (1 mentions) Ni nta agarose, by Qiagen (1 mentions) Salidroside, by MedChemExpress (1 mentions) Sb216763, by Apexbio (1 mentions) Pac 1, by BD (1 mentions) 7 aad, by Thermo Fisher Scientific (1 mentions) Matrine, by MedChemExpress (1 mentions) Fibrinogen, by Merck Group (1 mentions) Bsa bovine serum albumin, by Merck Group (1 mentions) Collagen and thrombin, by Chrono-Log (1 mentions)

8 protocols using pac 1 antibody

Integrin-Binding Protein Interaction Study

Cited 1 time

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

Flag antibody (SG4110–16, Shanghai Genomics), 6 × his antibody (SG4110–06, Shanghai Genomics), sharpin antibody (ab174545 and ab69507, Abcam), FAK antibody (#3283, CST) and Y-FAK antibodies (#3285, CST) were used for immunoblotting; PAC1 antibody (340535, BD Biosciences), 9EG7 antibody (553715, BD Biosciences) and 7E2 antibody (DSHB) were used for FACS analysis. Plasmid of GST-fibronectin type III repeats 9–11 (GST-Fn-III) was kindly provided by David Calderwood [18 (link)]. The cDNA of full length sharpin was kindly provide by Ivan Dikic [26 (link)], and subcloned into vectors of pET28a, pHis-1, pGST-1 and pGADT7 for different experiments. The CT of integrin α5β1 and integrin αIIbβ3 were subcloned into pGST-1 vector. Kindlin-1 was subcloned into pET31b, pGST-1 and pGBKT7 vectors. To express and purify proteins, the expression vectors were transformed into Rosetta DE3 strain and induced to express proteins with 0.4 mM of IPTG. GST-tagged or his-tagged proteins were purified by Glutathione Sepharpose (GE) and Ni-NTA agarose (Qiagen) respectively, according to the manufactures’ protocols. The purified GST-Fn-III protein was further labeled with biotin (EZ-Link™ NHS-Biotin, Thermo Fisher).

Gao J., Bao Y., Ge S., Sun P., Sun J., Liu J., Chen F., Han L., Cao Z., Qin J., White G.C., Xu Z, & Ma Y.Q. (2019). Sharpin suppresses β1-integrin activation by complexing with the β1 tail and kindlin-1. Cell Communication and Signaling : CCS, 17, 101.

+ Open protocol

+ Expand

Multiparametric Analysis of Platelet Activation

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

We used an LSR II flow cytometer (Beckon Dickinson, San Jose, CA, USA) to detect platelet activation through several measurements: CD62p expression, the binding of PAC-1 antibody (BD Biosciences), which recognizes the active conformation of the fibrinogen receptor αIIbβ3, annexin V binding,^{9 (link)} platelet-bound fibrinogen and coagulation factor V,^{25 (link),26 (link)} and the formation of platelet-leukocyte complexes (Online Supplementary Methods).

Zhao Z., Zhou Y., Hilton T., Li F., Han C., Liu L., Yuan H., Li Y., Xu X., Wu X., Zhang F., Thiagarajan P., Cap A., Shi F.D., Zhang J, & Dong J.F. (2020). Extracellular mitochondria released from traumatized brains induced platelet procoagulant activity. Haematologica, 105(1), 209-217.

+ Open protocol

+ Expand

Salidroside Modulates Platelet Activation

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

Salidroside was purchased from MedChemExpress (Monmouth Junction, NJ, USA) with a purity ≥ 98%. Collagen-related peptide (CRP) was prepared as previously described [56 (link)]. Thrombin (≥ 10 NIH units/vial) were from Chrono-log Corporation (Havertown, PA, USA). FITC-conjugated mouse anti-human CD41a and PAC-1 antibody were from BD Biosciences (San Jose, CA, USA) and BECTON DICKINSON (San Jose, CA, USA) respectively. PE-conjugated anti-human/mouse CD62p (P-Selectin) and anti-human Glycoprotein VI purified antibody were purchased from eBioscience (San Diego, CA, USA). FITC-conjugated anti-CD42b antibody was from Abcam (Cambridge, MA, USA). FITC-conjugated goat anti-mouse IgG was purchased from ZSGB-BIO (Beijing, China). β-actin antibody and anti-rabbit IgG (HRP-linked) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). SB216763 (GSK-3β inhibitor) was purchased from ApexBio Technology.

Wei G., Xu X., Tong H., Wang X., Chen Y., Ding Y., Zhang S., Ju W., Fu C., Li Z., Zeng L., Xu K, & Qiao J. (2020). Salidroside inhibits platelet function and thrombus formation through AKT/GSK3β signaling pathway. Aging (Albany NY), 12(9), 8151-8166.

+ Open protocol

+ Expand

Measuring αIIbβ3 Integrin Activation in CHO Cells

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

The αIIbβ3 integrin activation was assessed in CHO cells stably expressing human αIIbβ3 integrin (A5 cells)^{53 (link)} by flow cytometry using the ligand-mimetic antibody PAC1 (BD)^{54 (link)}. CHO cells were lipofected with 1 µg of plasmids expressing eGFP-tagged Kank2 constructs and/or 3 µg TagRFP-tagged talin-1 or GFP-tagged THD. Cells were trypsinized 24 h after transfection and incubated with the PAC1 antibody (BD) in Tyrode’s buffer (pH 7.35) for 30 min on ice, washed and stained with a secondary IgM for 30 min on ice followed by a final wash and staining with a streptavidin-Cy5 for 30 min on ice. PAC1 binding was measured with a FACS Canto, gated for living cells, using 7AAD (Thermo) exclusion as well as GFP–RFP double-positive cells. Total αIIbβ3 integrin surface levels were determined with an anti-αIIb integrin antibody (HIP8, BioLegend). Data evaluation was carried out with the FlowJo software. Surface antigen binding was expressed as geometric mean fluorescence intensity. αIIbβ3 integrin activation levels were defined as the ratio of PAC1 and HIP8 binding in cells positive for eGFP and/or RFP and were normalized with that of the cell expressing THD only.

Sun Z., Tseng H.Y., Tan S., Senger F., Kurzawa L., Dedden D., Mizuno N., Wasik A.A., Thery M., Dunn A.R, & Fässler R. (2016). Kank2 activates talin, reduces force transduction across integrins and induces central adhesion formation. Nature cell biology, 18(9), 941-953.

+ Open protocol

+ Expand

Platelet Activation Assay with Matrine

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

Matrine was purchased from MedChemExpress (Monmouth Junction, NJ, United States) with a purity ≥98% and dissolved in saline. Collagen-related peptide (CRP) was prepared as previously described (Arthur et al., 2012 (link)). Collagen and thrombin (≥10 NIH units/vial) were from Chrono-log Corporation (Havertown, PA, United States). Fibrinogen and BSA (bovine serum albumin) were purchased from Sigma-Aldrich (St. Louis, MO, United States). Mepacrine (also known as Quinacrine) was from APExBIO (Boston, MA, United States). FITC-conjugated mouse anti-human CD41a and PAC-1 antibody were from BD Biosciences (San Jose, CA, United States). PE-labelled anti-human/mouse CD62p (P-Selectin) and anti-human Glycoprotein VI antibody were purchased from eBioscience (San Diego, CA, United States). Alexa Fluor-546-labelled phalloidin was purchased from Thermo Fisher Scientific (Waltham, MA, United States). FITC-conjugated goat anti-mouse IgG was purchased from ZSGB-BIO (Beijing, China). β-actin antibody and HRP-conjugated anti-rabbit IgG were purchased from Cell Signaling Technology (Danvers, MA, United States).

Zhang S., Gui X., Ding Y., Tong H., Ju W., Li Y., Li Z., Zeng L., Xu K, & Qiao J. (2021). Matrine Impairs Platelet Function and Thrombosis and Inhibits ROS Production. Frontiers in Pharmacology, 12, 717725.

+ Open protocol

+ Expand

Platelet activation and degranulation

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

Platelets (10×10⁶/mL) were incubated with TRAP (40 mM) for 30 min at 37°C, and fixed with 4% PFA for 20 min at RT. The expression of CD62P (CLB-Thromb/6, Beckman Coulter) and CD63 (CLB-Gran/12, Beckman Coulter) was determined by flow cytometry (FC) as well as the conformational changes of glycoproteins (GPs) IIb/IIIa complex by PAC-1-antibody (BD Bioscience).

Marini I., Aurich K., Jouni R., Nowak-Harnau S., Hartwich O., Greinacher A., Thiele T, & Bakchoul T. (2019). Cold storage of platelets in additive solution: the impact of residual plasma in apheresis platelet concentrates. Haematologica, 104(1), 207-214.

+ Open protocol

+ Expand

Platelet Aggregation Inhibition by Platycodin D

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

Platycodin D (PD) was purchased from Fluka (Steinheim, Germany) with a purity ≥ 99%. Collagen, adenosine diphosphate (ADP), arachidonic acid and epinephrine were from Helena laboratories (Beaumont, Texas, USA). Thrombin was purchased from Sigma-Aldrich (St. Louis, MO, USA). FITC-conjugated mouse anti-human CD41a and PAC-1 antibody were from BD Biosciences (San Jose, CA, USA) and Becton–Dickinson (San Jose, CA, USA) respectively. PE-conjugated anti-human/mouse CD62p (P-Selectin) and anti-human Glycoprotein VI purified antibody were purchased from eBioscience (San Diego, CA, USA). FITC-conjugated anti-CD42b antibody was from Abcam (Cambridge, MA, USA). FITC-conjugated goat anti-mouse IgG was purchased from ZSGB-BIO (Beijing, China). β-actin antibody and anti-rabbit IgG (HRP-linked) antibody were purchased from Cell Signaling Technology (Danvers, MA, USA).

Luo Q., Wei G., Wu X., Tang K., Xu M., Wu Y., Liu Y., Li X., Sun Z., Ju W., Qi K., Chen C., Yan Z., Cheng H., Zhu F., Li Z., Zeng L., Xu K, & Qiao J. (2018). Platycodin D inhibits platelet function and thrombus formation through inducing internalization of platelet glycoprotein receptors. Journal of Translational Medicine, 16, 311.

+ Open protocol

+ Expand

Characterization of Medicinal XML Compound

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

XML used in our research was provided by Yunnan Tengchong Pharmaceutical Corporation (Yunnan, China), in accordance with applicable Good Manufacturing Practice (GMP). We performed HPLC analysis to identify the chemical characteristics of XML samples satisfying the national standards for XML (HPLC≥99%; Figure 2). Collagen, thrombin, and arachidonic acid (AA) were purchased from Chrono-Log Corp. (Havertown, PA, USA). Antibodies against total-Syk, phospho-Syk (Tyr525/526), total-PLCγ2, phospho-PLCγ2 (Tyr759), total-Akt, phospho-Akt (Thr308), total-GSK3β, phospho-GSK3β (Ser9), total-Erk1/2, phospho-Erk1/2 (Thr202/Tyr204), total-JNK, phospho-JNK (Thr183/Tyr185), total-p38, and phospho-p38 (Thr180/Tyr182) were obtained from cell signaling (Beverly, MA, USA). CD61 and CD62P (P-selectin) antibodies and PAC-1 antibody were purchased from BD Biosciences (San Jose, CA, USA). ECL Western blotting detection reagent was obtained from Pierce Chemical Co. (Rockford, Illinois, USA). XML was dissolved in saline solution and stored at room temperature until used.

Wang H., Ye Y., Wan W., Wang L., Li R., Li L., Yang L., Yang L., Gu Y., Dong L, & Meng Z. (2019). Xinmailong Modulates Platelet Function and Inhibits Thrombus Formation via the Platelet αIIbβ3-Mediated Signaling Pathway. Frontiers in Pharmacology, 10, 923.

+ 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!