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  4. HSP70/DnaK Protein, E. coli (P.pastoris, His)

The HSP70/DnaK protein is a molecular chaperone that primarily responds to stress, especially heat shock. It plays a crucial role in helping proteins fold correctly and prevent misfolding or aggregation during cellular stress. HSP70/DnaK Protein, E. coli (P.pastoris, His) is the recombinant E. coli-derived HSP70/DnaK protein, expressed by P. pastoris , with N-6*His labeled tag.

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Description

The HSP70/DnaK protein is a molecular chaperone that primarily responds to stress, especially heat shock. It plays a crucial role in helping proteins fold correctly and prevent misfolding or aggregation during cellular stress. HSP70/DnaK Protein, E. coli (P.pastoris, His) is the recombinant E. coli-derived HSP70/DnaK protein, expressed by P. pastoris , with N-6*His labeled tag.
Background

HSP70/DnaK protein functions as a molecular chaperone, primarily responding to stress conditions, such as heat shock. In times of cellular stress, this protein plays a crucial role in assisting the proper folding of other proteins, preventing misfolding or aggregation. The chaperone activity of HSP70/DnaK is especially vital for cellular homeostasis and survival under adverse environmental conditions, where maintaining the correct conformation of proteins becomes challenging. The protein's ability to recognize and assist in the refolding of misfolded or denatured proteins is essential for cellular resilience and the prevention of proteotoxicity induced by various stressors, particularly elevated temperatures.
Species

E.coli
Source

P. pastoris
Tag

N-6*His
Accession

Q1RGI8 (M1-K638)
Gene ID

89519394
Molecular Construction
N-term
6*His
HSP70 (M1-K638)
Accession # Q1RGI8
C-term
Synonyms
dnaK; HSP70; Heat shock 70kDa protein; Heat shock protein 70
AA Sequence

MGKIIGIDLGTTNSCVAIMDGTTPRVLENAEGDRTTPSIIAYTQDGETLVGQPAKRQAVTNPQNTLFAIKRLIGRRFQDEEVQRDVSIMPFKIIAADNGDAWVEVKGQKMAPPQISAEVLKKMKKTAEDYLGEPVTEAVITVPAYFNDAQRQATKDAGRIAGLEVKRIINEPTAAALAYGLDKGTGNRTIAVYDLGGGTFDISIIEIDEVDGEKTFEVLATNGDTHLGGEDFDSRLINYLVEEFKKDQGIDLRNDPLAMQRLKEAAEKAKIELSSAQQTDVNLPYITADATGPKHMNIKVTRAKLESLVEDLVNRSIEPLKVALQDAGLSVSDIDDVILVGGQTRMPMVQKKVAEFFGKEPRKDVNPDEAVAIGAAVQGGVLTGDVKDVLLLDVTPLSLGIETMGGVMTTLIAKNTTIPTKHSQVFSTAEDNQSAVTIHVLQGERKRAADNKSLGQFNLDGINPAPRGMPQIEVTFDIDADGILHVSAKDKNSGKEQKITIKASSGLNEDEIQKMVRDAEANAEADRKFEELVQTRNQGDHLLHSTRKQVEEAGDKLPADDKTAIESALTALETALKGEDKAAIEAKMQELAQVSQKLMEIAQQQHAQQQTAGADASANNAKDDDVVDAEFEEVKDKK

Molecular Weight

Approximately 71.1 kDa
Purity

Greater than 90% as determined by reducing SDS-PAGE.
Appearance

Lyophilized powder.
Formulation

Lyophilized after extensive dialysis against solution in 20 mM Tris-HC1, 0.5 M NaCl, 6% Trehalose, pH 8.0.
Endotoxin Level

<1 EU/μg, determined by LAL method.
Reconstitution

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH2O.
Storage & Stability

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.
Shipping

Room temperature in continental US; may vary elsewhere.
Documentation
Data Sheet (239 KB) SDS (251 KB)

COA (206 KB)

Handling Instructions (2659 KB)

HSP70/DnaK Protein, E. coli (P.pastoris, His) Related Classifications

Help & FAQs
  • How should lyophilized recombinant proteins be reconstituted and stored?

    1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss. 2. Taking 10 μg as an example, first add 20 μL of reconstituted solution provided by MCE and use a pipette to gently resuspend the lyophilized protein until it is fully dissolved.. (For most proteins, the reconstitution solution we provide is sterile water. If a diluent other than water is required, it will be indicated in the product's Certificate of Analysis (COA).). 3. Add an additional 80 μL of buffer/culture medium containing carrier protein (either 0.1% BSA, 5% HSA, 10% FBS, or 5% trehalose), and then use a pipette to gently mix until uniform. The final concentration is should not be lower than 100 μg/mL. 4. Aliquot at least 20 μL per tube. 5. After aliquoting, store it frozen at a temperature ranging from -20ºC to -80ºC, and it can be preserved for 3 to 6 months.

  • How should solution-form recombinant proteins be stored?

    1. The product can be stored in its original form and diluted as needed upon use. 2. Alternatively, dilute with a buffer/culture medium containing a carrier protein (either 0.1% BSA, 5% HSA, 10% FBS, or 5% alginate), mix well by pipetting, and ensure that

  • Why is it necessary to add carrier proteins?

    Carrier proteins are commonly added to enhance the stability of recombinant proteins, preventing them from adhering to the walls of the container during freezing or thawing processes. Plastic tubes have a certain adsorptive capacity for proteins, which may lead to difficulty in separating the protein from the tube walls, resulting in a decrease in the actual concentration of the protein in the solution and thus affecting its activity. To minimize such losses, it is recommended to add a commonly used carrier protein solution prior to the long-term storage of recombinant protein products.

  • Carrier protein types and options?

    In cases where the carrier protein is not expected to influence the experimental outcomes, an appropriate carrier protein, such as 0.1% BSA (Bovine Serum Albumin), 5% HSA (Human Serum Albumin), 10% FBS (Fetal Bovine Serum), or 5% trehalose, can be incorpo

  • Reconstitution Calculator

  • Dilution Calculator

  • Specific Activity Calculator

The reconstitution calculator equation

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration

Volume (to add to vial) = Mass (in vial) ÷ Desired Reconstitution Concentration
= ÷

The dilution calculator equation

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
× = ×
C1   V1   C2   V2

The specific activity calculator equation

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)
Unit/mg = 106 ÷ ng/mL

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HSP70/DnaK Protein, E. coli (P.pastoris, His)
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