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ReadyBlot Kidney Protein explorer

The kidney is responsible for several homeostatic functions, including the regulation of extracellular fluid volume, K+ balance, acid-base homeostasis and regulation of the tonicity of the body fluid. To perform these homeostatic functions, the kidneys regulate NaCl reabsorption, K+ secretion, net acid excretion, and water transport. The presence, the distribution and the localization of highly specialized ion or water transport pathways along different segments of the nephron are fundamental to the response of kidney to changes in water or solutes intake.

Cortex: as a whole, is cup-shaped, has inverted margins, and surrounds the renal medulla. In addition to the Glomerulus, Cortex contains the following nephron segments: Proximal tubule (with S1, S2 and 50% of S3 segments), 10 to 20% of Cortical Thick ascending limb (in the superficial cortex), distal tubule, and cortical collecting duct.

Outer Medulla: can be divided into 2 regions: Outer Stripe linked to the cortex, and Inner Stripe far from Cortex, and linked to the inner medulla. Inner Stripe of Outer medulla, which contain 100% of medullary thick ascending limb (mTAL) tubules, Thin descending limb, and Outer Medullary collecting duct (OMCD). Inner Medulla or the papilla projects into the renal pelvis. The papilla contains both Inner medullary collecting duct (IMCD) and the thin descending and ascending limb of Henle’s loop. Majority (70-80%) of water, electrolytes (Na+, Cl-, K+, HCO3-, Pi, etc) and organic solutes (Glucose, Amino acid, etc) filtered by the glomerulus are reabsorbed by the proximal tubules cells (localized in the cortex).

BLM and BBM are prepared from kidney cortex by Percoll density gradient method.

Acquisition of animal or human kidney tissue is not only time-consuming and expensive, but also requires expertise and training in kidney anatomy, cell and molecular biology. ADI has carefully dissected and processed anatomically and functionally distinct areas of kidney for the study of proteins using Western blots. The kidney proteins have been electrophoresed, electro-blotted, and blocked. A lane of pre-stained mol. wt markers is included in each blot to assist you in identifying the size of the proteins.

Each Kidney ReadyBlot has the proteins from the following regions:

Protein Load: ~20 ug protein; Further optimized for equal protein load (fig 2) and with beta-actin (fig. 3) immuno blot.

The proteins representing the total extracts of different regions of the kidney were stained with comassie (Fig. 2) or probed with beta-actin antibody (Fig. 3).

SDS-Gel Electrophoresis and blotting: Kidney protein extracts were mixed with 2X standard Laemmeli reducing buffer, heated for 5 min at 90oC. Approx 20 ug total proteins were run on 4-20%-reducing SDS-mini gels at 200 V for approx. 45 min. Pre-stained high range mol. Wt markers (ADI Cat # HMWM-11) were loaded on each gel:

The proteins were transferred to nitrocellulose or PVDF using mini-transblot cells. Homogeneity of protein transfer in all 12 lanes was verified using water soluble Stain-ALL (ADI Cat # SALL-500) for 5 min. Protein lanes were identified and marked 1-8. Membranes were washed in PBS to remove the dye. Multi-colored mol. Wt standards have been marked A-H on the blot.

Blocking: After destaining, membranes were blocked with 1:10 diluted PBS/milk-based buffer (ADI Cat# 80062) and dried for proper storage.

Form, Storage, and Recommended Usage: Blots are provided pre-blocked and in ready-to-use forms. Store unused blots at 4oC in a sealed bag. ReadyBlot should be handled with care as blotting membrane is quiet brittle. These blots should be used within 3-4 months. It is recommended to wet the blot first in appropriate buffer before incubating with antibodies.

This blot will be most useful for proteins that are relatively abundant in whole kidney tissue. Very low abundant proteins that require the use of enriched cell membranes or nuclear fractions may be poorly represented in whole tissue blots. .

Adult Mouse ReadyBlot Kidney Protein Explorer
Cat # MKWB-61; $495.00 (Mouse: Swiss Webster, ~10 wks old, mixed gender)

Adult Rat ReadyBlot Kidney Protein Explorer
Cat # RKWB-81; $495.00 (Rat: Sprague-Dawley, ~ 8 wks old, mixed gender)

ReadyBlots Protein Explorer from the following tissues are also available:

Western Blot Recycling Kit (Reuse ReadyBlot or any other blot many times by stripping antibodies at room temp.

All Products are for in vitro research use only.