Image 1: Structure of the albumin target protein.

Albumin target introduction

Protein function

• Albumin is the most abundant circulating protein in plasma, accounting for half of the total protein content in a normal human body (3.5-5 g/dL).
• Albumin can regulate plasma osmotic pressure, transport endogenous (such as hormones, fatty acids, metabolic products) and exogenous (such as drugs) ligands in the body. This protein also exhibits broad substrate-specific esterase activity.
• Albumin is synthesized by hepatocytes and stored in the liver in small amounts, most of which are rapidly released into the blood at a rate of 10-15 gm/d.
• Albumin is a colloid fluid that can be used for fluid resuscitation in patients, especially in cases of trauma (such as hypovolemic shock) or large-volume abdominal puncture. Albumin can reflect the patient's liver function status or the ability to synthesize proteins, and is an important factor in maintaining self-stability.

Protein expression

• Plasma

Protein localization

• Secretion

Figure 2: ICC experimental result image of Albumin protein, using Anti-Albumin antibody (ab207327).

Green: Albumin, Red: alpha Tubulin, Blue: DAPI.

Isoforms & Post-translational modifications

• Human (P02768):
  Isoform 1 (P02768-1): 69.3 kDa (predicted)
  Isoform 2 (P02768-2): 47.3 kDa (predicted)
  Isoform 3 (P02768-3): 45.1 kDa (predicted)
• Mouse (P07724): 68.6 kDa (predicted)
• Rat (P02770): 68.7 kDa (predicted)
• Glycosylation
• Phosphorylation
• Acetylation

WB experiment tips

Precautions

• Please note that the expression level of Albumin may vary in different samples, and it is necessary to confirm the expression level of the target protein before testing. It is recommended to use the samples mentioned in the antibody product manual as positive controls.

Positive controls

• Human liver tissue lysate, plasma lysate
• Rat liver tissue lysate, plasma lysat
• Mouse liver tissue lysate, plasma lysate

Negative control (no expression or weak expression)

• Human HeLa cell lysate

Example of results

Figure 3: Western blot results of Albumin protein, Anti-Albumin antibody (ab207327).

Lane 1: Human liver tissue lysate (20ug).
Lane 2: Mouse liver tissue lysate (20ug).
Lane 3: Rat liver tissue lysate (20ug).
Lane 4: HepG2 whole cell lysate (20ug).
Lane 5: HeLa whole cell lysate (10ug).
Lanes 1-3: Exposure for 5 seconds.
Lanes 4/5: Exposure for 1 second.

Predicted band size: 69 kDa
Detected band size: 69 kDa

Key control points

In the experiment, in addition to paying attention to routine issues, special attention should be paid to the following key control points:

Sample preparation:

1. Add a complex protease inhibitor to avoid degradation of the target protein.
2. Select a suitable lysis buffer to enrich more target proteins.
3. Sonicate the cells to enrich the target protein.
4. Keep the sample on ice throughout the sample preparation process.
5. Determine the protein concentration of the sample through Bradford analysis, Lowry analysis, or BCA analysis.

Blocking:

1. There is no blocking solution that is suitable for all systems; we recommend choosing the appropriate blocking solution for your experiment

Antibody incubation:

1. In the WB experiment, please avoid dry film conditions.
2. Please choose the appropriate antibody working concentration according to the product manual.
3. It is recommended to use fresh antibodies and not to reuse antibodies.

Reference

1. Gregory J. Quinlan, Greg S. Martin, Timothy W. Albumin: Biochemical properties and therapeutic. Hepatology. (2005) 41(6), 1211-1219. doi: https://doi.org/10.1002/hep.20720
2. Yuwen P, Chen W, Lv H et al. Albumin and surgical site infection risk in orthopaedics: a meta-analysis. BMC Surg. (2017) 17(1),7. doi: https://doi.org/10.1186/s12893-016-0186-6
3. MA Rothschild, M Oratz, SS Schreiber. Serum albumin. Hepatology. (1988) 8(2), 385-401. doi: https://doi.org/10.1002/hep.1840080234
4. Annane D, Siami S, Jaber S et al. Effects of fluid resuscitation with colloids vs crystalloids on mortality in critically ill patients presenting with hypovolemic shock: the CRISTAL randomized trial. JAMA. (2013) 310(17):1809-1817. doi: http://doi.org/10.1001/jama.2013.280502