Log In Sign up
The largest database of trusted experimental protocols

Fitc conjugated pac1 antibody

Manufactured by BD
Visit Supplier

The FITC-conjugated PAC1 antibody is a fluorescently labeled monoclonal antibody that specifically binds to the PAC1 receptor. PAC1 is a G protein-coupled receptor that plays a role in various physiological processes. The FITC (Fluorescein Isothiocyanate) label allows for the detection and visualization of PAC1-expressing cells using fluorescence-based techniques.

Automatically generated - may contain errors

Lab products found in correlation

Actin, by Cell Signaling Technology (2 mentions) P s6kt389, by Cell Signaling Technology (2 mentions) Total s6k, by Cell Signaling Technology (2 mentions) Cd41 fitc, by BioLegend (2 mentions) Annexin 5 fitc, by BioLegend (2 mentions) Rat insulin elisa kit, by ALPCO (2 mentions) Pakt t308, by Cell Signaling Technology (2 mentions) Human rcela2a, by MyBioSource (2 mentions) Cela2a elisa kit, by MyBioSource (2 mentions) Cela2a antibody, by Merck Group (2 mentions) N succinyl ala ala pro leu p nitroanilide substrate, by Merck Group (2 mentions) Tetramethylrhodamine ethyl ester tmre reagent, by Thermo Fisher Scientific (2 mentions) Total irs, by Cell Signaling Technology (2 mentions) Pirs y608, by Cell Signaling Technology (2 mentions) Pirs s636 s639, by Cell Signaling Technology (2 mentions) A1at antibody ab9400, by Abcam (2 mentions) Itga2b antibodies, by Abcam (2 mentions) Human c peptide elisa kit, by ALPCO (2 mentions) Human glucagon elisa kit, by Mercodia (2 mentions) Rat c peptide elisa, by ALPCO (2 mentions)

8 protocols using fitc conjugated pac1 antibody

1

Quantitative Platelet and Pancreatic Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified GP2B3A complex (MBS135714), mouse rCela2a protein (MBS1246487), human rCELA2A (MBS1090462) and the CELA2A ELISA kit (MBS932150) were purchased from MyBiosource. CELA2A antibody (SAB1104798) and the elastase substrate N-Succinyl-Ala-Ala-Pro-Leu p-nitroanilide substrate (S8511) were purchased from Sigma-Aldrich. The FITC-conjugated PAC1 antibody (340507, BD Biosciences) and tetramethylrhodamine ethyl ester (TMRE) reagent (T669, Thermo Fisher Scientific), Annexin V-FITC (640905, Biolegend), CD41-FITC (303703, Biolegend) were used for platelet aggregation analysis. Total AKT (CST, 9272), pAKT (S473) (CST, 9271s), pAKT (T308) (CST, 9275s), total S6k (CST, 9202), pS6k (T389) (CST, 9206), total IRS (CST, 2382), pIRS (Y608), (CST, 2385S), pIRS (S636/S639) (CST, 2388s), Gapdh (CST, 3683s), and Actin (CST, 4970) antibodies were purchased from Cell Signaling Technologies. Recombinant A1AT (alpha1 antitrypsin, ab91136), A1AT antibody (ab9400) and ITGA2B antibodies (ab63983) were purchased from Abcam. Human c-peptide ELISA kit (80-CPTHU-E01.1, ALPCO), Human Glucagon ELISA kit (10-1271-01, Mercodia), Rat c-peptide ELISA (80-CPTRT-E01, ALPCO), Mouse Ultrasensitive Insulin ELISA (80-INSMSU-E01, ALPCO), Human Insulin ELISA kit (21-IAAHU-E01, ALPCO), Rat Insulin ELISA kit (80-INSRT-E01, ALPCO) was applied for C-peptide, glucagon, and insulin measurements.
Esteghamat F., Broughton J.S., Smith E., Cardone R., Tyagi T., Guerra M., Szabó A., Ugwu N., Mani M.V., Azari B., Kayingo G., Chung S., Fathzadeh M., Weiss E., Bender J., Mane S., Lifton R.P., Adeniran A., Nathanson M.H., Gorelick F.S., Hwa J., Sahin-Tóth M., Belfort-DeAguiar R., Kibbey R.G, & Mani A. (2019). CELA2A mutations predispose to early-onset atherosclerosis and metabolic syndrome and affect plasma insulin and platelet activation. Nature genetics, 51(8), 1233-1243.
+ Open protocol
+ Expand
2

Platelet Activation and Apoptosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The PAC1 assay was used to analyze activation of GP2B3A receptor on platelet surface as described44 . This assay uses an antibody that binds specifically to activated conformation of human GP2B3A receptor and not to its resting form44 . The pretreated washed platelets were used for this assay. The FITC conjugated PAC1 antibody (340507, BD Biosciences) was added to washed platelets and after 20 min incubation before acquiring the events on flow cytometer. For apoptosis assay, the pretreated washed platelets were incubated with 1 μM tetramethylrhodamine methyl ester (TMRE) reagent (T669, Thermo Fisher Scientific) at 37 °C for 15 min followed by staining with annexin-V for 15 min in apoptosis assay buffer, as described45 . This assay quantifies the cells with depolarized mitochondrial membrane and exposed phosphatidyl serine on the platelet surface. For both the assays, 3 × 104 events were acquired using special order BD LSR II Cell Analyzer (BD Biosciences) and data were analyzed by FlowJo v10 (FlowJo LLC).
Esteghamat F., Broughton J.S., Smith E., Cardone R., Tyagi T., Guerra M., Szabó A., Ugwu N., Mani M.V., Azari B., Kayingo G., Chung S., Fathzadeh M., Weiss E., Bender J., Mane S., Lifton R.P., Adeniran A., Nathanson M.H., Gorelick F.S., Hwa J., Sahin-Tóth M., Belfort-DeAguiar R., Kibbey R.G, & Mani A. (2019). CELA2A mutations predispose to early-onset atherosclerosis and metabolic syndrome and affect plasma insulin and platelet activation. Nature genetics, 51(8), 1233-1243.
+ Open protocol
+ Expand
3

Monocyte-Platelet Aggregation and Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
As previously described, 14 (link) monocyte platelet aggregation (MPA) was measured via whole blood flow cytometry using the Accuri 6 system. Immediately following phlebotomy, whole blood was fixed with formaldehyde in 1.4× Hanks balanced saline solution. Blood was labeled with monoclonal antibodies CD14-PE (BD Pharmingen) and CD42a-FITC (BD Pharmingen); diluted 4.6-fold with distilled water to lyse the erythrocytes; and further diluted in an equal volume of HEPES–Tyrode’s buffer, pH 7.4. Monocytes were identified by their staining with CD14-PE and by their characteristic orthogonal light scatter. The percentage of MPA was identified in single parameter histograms of CD42a-FITC fluorescence displaying events from the monocyte gate. The positive analysis region was determined using an IgG-FITC conjugated isotypic control. A minimum of 2,000 monocytes was counted per test.
Platelet activity was also assessed by activation of the platelet integrin αIIbβ3 activity using a FITC-conjugated PAC-1 antibody (BD Biosciences). Platelets were labeled with anti CD42b PE antibody (BD Biosciences). The mean fluorescent intensity (MFI) was assessed within a participant gated sample of approximately 10,000 platelets.
Schneider G.S., Rockman C.B, & Berger J.S. (2014). Platelet Activation Increases in Patients Undergoing Vascular Surgery. Thrombosis research, 134(5), 952-956.
+ Open protocol
+ Expand
4

Platelet Activation Assay by PAC1

Check if the same lab product or an alternative is used in the 5 most similar protocols
For human platelets, the PAC1 assay was used to analyze activation of αIIbβ3 receptor on platelet surface. The PAC1 antibody binds specifically to the activated conformation of human αIIbβ3 receptor and not to its resting form [17 (link)].The FITC conjugated PAC1 antibody (BD Biosciences) was added to washed platelets and after 20 min incubation the events were acquired on LSRII. The absence of contaminating WBCs in platelet population was further confirmed at this step.
Jain K., Tyagi T., Patell K., Xie Y., Kadado A.J., Lee S.H., Yarovinsky T., Du J., Hwang J., Martin K.A., Testani J., Ionescu C.N, & Hwa J. (2019). Age associated non-linear regulation of redox homeostasis in the anucleate platelet: Implications for CVD risk patients. EBioMedicine, 44, 28-40.
+ Open protocol
+ Expand
5

Quantitative Platelet and Pancreatic Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Purified GP2B3A complex (MBS135714), mouse rCela2a protein (MBS1246487), human rCELA2A (MBS1090462) and the CELA2A ELISA kit (MBS932150) were purchased from MyBiosource. CELA2A antibody (SAB1104798) and the elastase substrate N-Succinyl-Ala-Ala-Pro-Leu p-nitroanilide substrate (S8511) were purchased from Sigma-Aldrich. The FITC-conjugated PAC1 antibody (340507, BD Biosciences) and tetramethylrhodamine ethyl ester (TMRE) reagent (T669, Thermo Fisher Scientific), Annexin V-FITC (640905, Biolegend), CD41-FITC (303703, Biolegend) were used for platelet aggregation analysis. Total AKT (CST, 9272), pAKT (S473) (CST, 9271s), pAKT (T308) (CST, 9275s), total S6k (CST, 9202), pS6k (T389) (CST, 9206), total IRS (CST, 2382), pIRS (Y608), (CST, 2385S), pIRS (S636/S639) (CST, 2388s), Gapdh (CST, 3683s), and Actin (CST, 4970) antibodies were purchased from Cell Signaling Technologies. Recombinant A1AT (alpha1 antitrypsin, ab91136), A1AT antibody (ab9400) and ITGA2B antibodies (ab63983) were purchased from Abcam. Human c-peptide ELISA kit (80-CPTHU-E01.1, ALPCO), Human Glucagon ELISA kit (10-1271-01, Mercodia), Rat c-peptide ELISA (80-CPTRT-E01, ALPCO), Mouse Ultrasensitive Insulin ELISA (80-INSMSU-E01, ALPCO), Human Insulin ELISA kit (21-IAAHU-E01, ALPCO), Rat Insulin ELISA kit (80-INSRT-E01, ALPCO) was applied for C-peptide, glucagon, and insulin measurements.
Esteghamat F., Broughton J.S., Smith E., Cardone R., Tyagi T., Guerra M., Szabó A., Ugwu N., Mani M.V., Azari B., Kayingo G., Chung S., Fathzadeh M., Weiss E., Bender J., Mane S., Lifton R.P., Adeniran A., Nathanson M.H., Gorelick F.S., Hwa J., Sahin-Tóth M., Belfort-DeAguiar R., Kibbey R.G, & Mani A. (2019). CELA2A mutations predispose to early-onset atherosclerosis and metabolic syndrome and affect plasma insulin and platelet activation. Nature genetics, 51(8), 1233-1243.
+ Open protocol
+ Expand
6

Peptide HNG-induced Platelet Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Peptide HNG was synthesized from Shanghai Bootech Bioscience and Technology Co., Ltd. Anti‐acetylated tubulin antibody (T6793) and anti‐β1 tubulin antibody (ab179511) were purchased from Sigma‐Aldrich and Abcam. Anti‐phospho‐PLCγ2 (Y1217), anti‐phospho‐ERK1/2 (Thr202/Tyr204), anti‐phospho‐p38 MAPK (Thr180/Tyr182), anti‐phospho‐Akt (Ser473), anti‐PLCγ2, anti‐ERK1/2, anti‐p38 MAPK and anti‐Akt antibodies were purchased from Cell Signaling Technologies. Phalloidin‐TRITC was obtained from Sigma‐Aldrich. PE‐conjugated mouse anti‐human CD62P antibody and FITC‐conjugated PAC‐1 antibody were purchased from BD Biosciences.
Ren L., Li Q., You T., Zhao X., Xu X., Tang C, & Zhu L. (2020). Humanin analogue, HNG, inhibits platelet activation and thrombus formation by stabilizing platelet microtubules. Journal of Cellular and Molecular Medicine, 24(8), 4773-4783.
+ Open protocol
+ Expand
7

Platelet Activation and Apoptosis Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The PAC1 assay was used to analyze activation of GP2B3A receptor on platelet surface as described44 . This assay uses an antibody that binds specifically to activated conformation of human GP2B3A receptor and not to its resting form44 . The pretreated washed platelets were used for this assay. The FITC conjugated PAC1 antibody (340507, BD Biosciences) was added to washed platelets and after 20 min incubation before acquiring the events on flow cytometer. For apoptosis assay, the pretreated washed platelets were incubated with 1 μM tetramethylrhodamine methyl ester (TMRE) reagent (T669, Thermo Fisher Scientific) at 37 °C for 15 min followed by staining with annexin-V for 15 min in apoptosis assay buffer, as described45 . This assay quantifies the cells with depolarized mitochondrial membrane and exposed phosphatidyl serine on the platelet surface. For both the assays, 3 × 104 events were acquired using special order BD LSR II Cell Analyzer (BD Biosciences) and data were analyzed by FlowJo v10 (FlowJo LLC).
Esteghamat F., Broughton J.S., Smith E., Cardone R., Tyagi T., Guerra M., Szabó A., Ugwu N., Mani M.V., Azari B., Kayingo G., Chung S., Fathzadeh M., Weiss E., Bender J., Mane S., Lifton R.P., Adeniran A., Nathanson M.H., Gorelick F.S., Hwa J., Sahin-Tóth M., Belfort-DeAguiar R., Kibbey R.G, & Mani A. (2019). CELA2A mutations predispose to early-onset atherosclerosis and metabolic syndrome and affect plasma insulin and platelet activation. Nature genetics, 51(8), 1233-1243.
+ Open protocol
+ Expand
8

Activated αIIbβ3 Receptor Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
For human platelets, the PAC1 assay was used to analyze the activation of αIIbβ3 surface receptor on platelets. The PAC1 antibody binds specifically to the activated conformation of human αIIbβ3 receptor and not to its resting form (Shattil et al., 1985 (link)). The FITC-conjugated PAC1 antibody (BD Biosciences) was added to washed platelets as per the manufacturer’s instructions, and after 30 min of incubation at room temperature, the reaction was stopped and the events were acquired on an LSRII.
Tyagi T., Jain K., Yarovinsky T.O., Chiorazzi M., Du J., Castro C., Griffin J., Korde A., Martin K.A., Takyar S.S., Flavell R.A., Patel A.A, & Hwa J. (2022). Platelet-derived TLT-1 promotes tumor progression by suppressing CD8+ T cells. The Journal of Experimental Medicine, 220(1), e20212218.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!