Log In Sign up
The largest database of trusted experimental protocols

Clone 16b12

Manufactured by BioLegend
Visit Supplier

Clone 16B12 is a mouse monoclonal antibody that recognizes the human CD45 antigen. CD45 is a transmembrane protein tyrosine phosphatase that is expressed on the surface of all nucleated hematopoietic cells.

Automatically generated - may contain errors

Lab products found in correlation

Precision plus protein dual color standard, by Bio-Rad (1 mentions) Rabbit anti ha, by Merck Group (1 mentions) Irdye 680rd donkey anti rabbit, by LI COR (1 mentions) Image studio lite v5, by LI COR (1 mentions) Irdye 800cw goat anti mouse, by LI COR (1 mentions) Mouse anti flag, by Merck Group (1 mentions) Mouse anti ha, by BioLegend (1 mentions) H 215, by Santa Cruz Biotechnology (1 mentions) H 119, by Santa Cruz Biotechnology (1 mentions) Ab144p, by Merck Group (1 mentions) Clone ac 15, by Merck Group (1 mentions) Continuum transfection reagent, by Gemini Bio (1 mentions) Trueblot, by Rockland Immunochemicals (1 mentions) Dynabeadstm protein g, by Thermo Fisher Scientific (1 mentions)

5 protocols using clone 16b12

1

GPCR Immunoprecipitation and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunoprecipitation experiments, WTT-CHO cells were transiently transfected with pcDNA3.1 (+) empty vector (control) or vectors encoding HA-GPCRs in similar conditions to those used for AFM-SMFS experiments. Forty-eight hours post-transfection, cells were lysed in a lysis buffer (140 mM NaCl/2 mM EDTA/25 mM Tris, pH 7.4/0.5% DDM) supplemented with complete protease and phosphatase inhibitors (Roche). Protein extracts (~2 mg) were immunoprecipitated overnight at 4 °C with DynabeadsTM Protein G (Invitrogen) precoated with an anti-HA antibody (BioLegend, 16B12 Clone) and proceeded to western-blot analysis as previously described33 (link). Proteins were detected with the anti-HA primary antibodies (Biolegend, 16B12 Clone) followed by antimouse Horse Radish Peroxidase (HRP)-conjugated secondary antibodies (TrueBlot, eB144Clone, Rockland) using enhanced chemiluminescence detection reagent (RPN2232 Prime, GE Healthcare).
Dague E., Pons V., Roland A., Azaïs J.M., Arcucci S., Lachaize V., Velmont S., Trevisiol E., N’Guyen D., Sénard J.M, & Galés C. (2022). Atomic force microscopy-single-molecule force spectroscopy unveils GPCR cell surface architecture. Communications Biology, 5, 221.
+ Open protocol
+ Expand
2

Western Blotting of HA- and FLAG-tagged Proteins

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
We performed western blotting as in Polino et al. (2020) (link) with primary antibodies mouse anti-HA diluted 1:1000 (clone 16B12; Biolegend, 901501), rabbit anti-HA diluted 1:1000 (Sigma-Aldrich, H6908), rabbit anti-PfAldolase diluted 1:2000 (Abcam, ab207494; targets the protein with PlasmoDB accession PF3D7_1444800) and mouse anti-FLAG diluted 1:500 (Sigma-Aldrich, F1804), followed by secondary antibodies goat anti-mouse IRDye 800CW (Licor) and donkey anti-rabbit IRDye 680RD (Licor), both diluted 1:10,000. For Fig. 2C, parasites were harvested at the indicated times and Pf3D7_1437000-3xHA levels were quantified using ImageStudio Lite v. 5.2 (Licor). The sizes of bands were approximated using the Precision Plus Protein Dual Color Standards (Bio-Rad, 1610374).
Polino A.J., Hasan M.M., Floyd K., Avila-Cruz Y., Yang Y, & Goldberg D.E. (2023). An essential endoplasmic reticulum-resident N-acetyltransferase ortholog in Plasmodium falciparum. Journal of Cell Science, 136(6), jcs260551.
+ Open protocol
+ Expand
3

HA-Tagged Protein Immunoprecipitation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoprecipitation using mouse monoclonal anti-HA antibody (1:50; clone 16B12, BioLegend) were performed as previously reported with minor modification (37 (link)).
Guo X., Kimura A., Namekata K., Harada C., Arai N., Takeda K., Ichijo H, & Harada T. (2022). ASK1 signaling regulates phase-specific glial interactions during neuroinflammation. Proceedings of the National Academy of Sciences of the United States of America, 119(6), e2103812119.
+ Open protocol
+ Expand
4

Immunohistochemical Analysis of 5-HT1A Receptors

Check if the same lab product or an alternative is used in the 5 most similar protocols
The primary antibodies utilized were: rat anti-serotonin antibody (1:100 dilution, clone YC5/45, Merck), rabbit anti-ankyrin-G (anti-ankG; 1:300 dilution, H-215, Santa Cruz Biotechnology), mouse anti-microtubule-associated protein-2 (anti-MAP-2; 1:250 dilution, A-4, Santa Cruz Biotechnology), goat anti-choline acetyltransferase (anti-ChAT; 1:100 dilution. AB144P, Merck Life Science A/S), rabbit anti-5-HT1A (1:100, H-119, Santa Cruz Biotechnology), rabbit anti-5-HT1A (1:50, ASR-021, Alomone Labs), rabbit anti-5-HT1A (1:100, ADI905-741-100, Enzo Life Sciences), rabbit 5-HT1A (1:100, NB100-92418, Novus Biologicals), and mouse anti-HA epitope (1:500 dilution, clone 16B12, Biolegend). Rabbit 5HT1A S1A-170 serum (1:1000–1:2500 dilutions) was a generous gift from Professor Efrain C. Azmitia, New York University, New York, NY, USA.
Perrier J.F., Rasmussen H.B., Jørgensen L.K, & Berg R.W. (2018). Intense Activity of the Raphe Spinal Pathway Depresses Motor Activity via a Serotonin Dependent Mechanism. Frontiers in Neural Circuits, 11, 111.
+ Open protocol
+ Expand
5

Radioactive Labeling of Inositol Phosphates in HEK293A Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
HEK293A cells were plated at a density of ~75000 cells/well in 12-well tissue culture dishes in growth medium [DMEM containing 10% (v/v) FBS, 100 units/mL penicillin, 100 μg/mL streptomycin, and 0.25 μg/mL amphotericin B]. Twenty-four hours later, cells were transiently transfected with 100 ng of a plasmid encoding HA-tagged wild-type PLC-γ1 or PLC-γ1 (D1165H) using Continuum transfection reagent according to the manufacturer’s protocol (GeminiBio). Twenty-four hours post-transfection, growth medium was removed and cells were metabolically labeled overnight with 1 μCi of [3H]-myo-inositol in serum-free, inositol-free DMEM. Radiolabeling medium was then removed and replaced with vehicle [serum-free, inositol-free DMEM containing 1 μCi of [3H]-myo-inositol and 1% (v/v) DMSO] or a vehicle containing 30 μM Hit-1, and cells were incubated for 1 h. Cells were then treated with 10 mM LiCl (final concentration) for an additional 1 h, and accumulation of [3H]inositol phosphates was quantified by Dowex column chromatography as described previously. Expression of each version of PLC-γ1 was confirmed by immunoblotting of cell lysates using a monoclonal antibody against the HA epitope (BioLegend, clone 16B12; RRID: AB_2565335). A monoclonal antibody against β-actin (SigmaAldrich, clone AC-15; RRID: AB_476692) was used as a loading control.
Huang W., Carr A.J., Hajicek N., Sokolovski M., Siraliev-Perez E., Hardy P.B., Pearce K.H., Sondek J, & Zhang Q. (2020). A High-Throughput Assay to Identify Allosteric Inhibitors of the PLC-γ Isozymes Operating at Membranes. Biochemistry, 59(41), 4029-4038.
+ 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!