PsaC | PSI positive control/quantitation standard

PsaC | PSI positive control/quantitation standard

APA-PSQ-013

PsaC is a conserved, chloroplast-encoded, Fe-S binding protein of approximately 10kDa, present in all known Photosystem I complexes. It is located on the stromal side of the thylacoid membranes. PsaC coordinates the Fe–S clusters FA and FB through two cysteine-rich domains. This product is a recombinant protein standard, source: Synechocystis PCC 6803.The PsaC protein standard can be used in combination with global anti-PsaC antibodies to quantitate PsaC from a wide range of species. Global antibodies are raised against highly conserved amino acid sequences in the PsaC protein.Quantitative western blot:  detailed method description, video tutorial

Lyophilized in glycerol.

100 µl

For reconstitution add 95 µl of sterile water. Note that due to glycerol in buffer, the lyophilized product appears as a dense liquid rather than a powder. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently. Avoid vigorous vortexing, as buffer contains detergent. Upon reconstitution, this standard is ready-to-load and does not require any additions or heating. See additional Handling Instructions below. PsaC standard protein concentration: 0.10 pmol/µl.

11.5 kDa (larger than native protein due to the addition of His-tag), In most gels PsaC migrates between 9 and 14 kDa

Western blot (WB)

Positive control: a 2 μL load per well is optimal for most chemiluminescent detection systems. Standard curve: 3 loads are recommended (eg. 0.5, 2 and 4μL). For most applications a sample load of 0.2 μg of chlorophyll will give a PsaC signal in this range. Exact loads can vary with the sensitivity of your system and the abundance of the target protein in your samples. Note: Optimal quantitation is achieved using moderate sample loads/well, generally 1 to 5 ug total protein. A trial experiment may be required i) to bring your sample load within the standard curve range and ii) to obtain a signal that is strong enough to reliably quantify but not so strong as to consume ECL reagents too quickly or saturate your detection system. These goals may achieved by adjusting both sample and standard loads.

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.