PsaC | PSI positive control/quantitation standard

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Cat: PA00987

Size: 100 µl

Datasheet:

Product Info

Format

Lyophilized in glycerol.

Reconstitution

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.

Storage

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.

Application

Western blot (WB)

Recommended dilution

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.

Expected | apparent MW

11,5 kDa (larger than native protein due to the addition of His-tag), In most gels PsaC migrates between 9 and 14 kDa

Reactivity

Additional information

Handling Instructions*IMPORTANT: In our experience, viscous liquids are surprisingly stable; insufficient mixing is the most common reason for unsatisfactory results. Following mixing, briefly pulse in a microcentrifuge to collect material from cap.Standard needs to be fully thawed and thoroughly mixed before each use. Proteins tend to stratify with the more dense layer after freezing. We recommend bringing the product to room temperature and either mixing by inverting or flicking tube 5-10 times. Pipetting up and down may also provide sufficient mixing, provided the tip is moved within the tube while taking up and expelling the liquid.,Protein standard buffer composition: Protein standard buffer composition: Glycerol 10%, Tris Base 141 mM, Tris HCl 106 mM, LDS 2%, EDTA 0.51 mM, SERVA Blue G250 0.22 mM, Phenol Red 0.175 mM, pH 8.5, 0.1mg/ml PefaBloc protease inhibitor (Roche), 50mM DTT.

Description

PsaC is a conserved, chloroplast-encoded, approximately 10kDa FE-S-binding protein present in all known photosystem I complexes. It is located on the stromal side of the thylakoid membrane. PsaC coordinates the Fe-S clusters FA and FB through two cysteine-rich domains. This product is the standard of recombinant protein, source: Polycytidis pcc6803. PsaC protein standards can be used in combination with global anti-PSAC antibodies to quantify PsaC in a variety of species. Global antibodies are proposed for highly conserved amino acid sequences in PsaC proteins. Quantitative Immunoblotting: Detailed method instructions, video tutorials

For research use only, not for clinical use.