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WNK Lysine Deficient Protein Kinase 1 (WNK1) (C-Term) antibody

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(24), (17), (16), (7), (7), (5), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Human, Mouse (Murine), Rat (Rattus)
(118), (44), (44), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1)
(118), (1)
(6), (4), (4), (4), (4), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2)
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF)
(58), (41), (40), (24), (22), (9), (5), (4), (1), (1), (1)
Pubmed 3 references available
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Quantity 0.1 mL
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Immunogen A synthetic peptide within the C-terminal region of human WNK1 (within the last 350 amino acids).
Purification affinity purified
Alternative Name WNK1 (WNK1 Antibody Abstract)
Background WNK kinases play an important role in hypertension. WNK1 is a cytoplasmic proteinthat localizes to a segment of kidney involved in salt, potassium, and pH homeostasis,the distal nephron. Alternate Names: anti-Kinase deficient protein antibody, anti-Protein kinase lysine deficient 1 antibody,anti-Protein kinase with no lysine 1 antibody, anti-Serine/threonine protein kinase WNK1antibody.
Gene Symbol: WNK1
Gene ID 65125, 116477
UniProt Q9H4A3
Research Area Signaling, Kinases/Phosphatases
Application Notes This WNK1 antibody is useful for Immunocytochemistry/Immunofluorescence and Western blot. In WB, this antibody recognizes the short form of WNK1, ~190 kDa. The theoretical molecular weight of human WNK1 is ~251 kDa and rat WNK1 is ~230 kDa. In ICC/IF cytomplasmic staining was observed.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:500 - 1:1000, Western Blot 1:1000
Protocol Western blot Protocol for WNK1 antibody
Western Blot Protocol:
1. Perform SDS-PAGE (4-12 % MOPS) on samples to be analyzed, loading 25 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % NFDM + 1 % BSA in TBS + Tween, 1 hour at RT.
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Dilute the rabbit anti-WNK1 primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions (Pierce ECL).Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.Immunocytochemistry/Immunofluorescence Protocol for WNK1 antibody Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Restrictions For Research Use only
Format Liquid
Concentration 5 mg/mL
Buffer PBS, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Avoid freeze-thaw cycles
Storage -20 °C
Storage Comment -20 °C.
Supplier Images
Western Blotting (WB) image for anti-WNK Lysine Deficient Protein Kinase 1 (WNK1) (C-Term) antibody (ABIN153167) anti-WNK Lysine Deficient Protein Kinase 1 (WNK1) (C-Term) antibody
Product cited in: Wade, Fang, Liu et al.: "WNK1 kinase isoform switch regulates renal potassium excretion." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 103, Issue 22, pp. 8558-63, 2006 (PubMed).

Nuaray-Fejes-Tuoth, Snyder, Fejes-Tuoth: "The kidney-specific WNK1 isoform is induced by aldosterone and stimulates epithelial sodium channel-mediated Na+ transport." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, Issue 50, pp. 17434-9, 2004 (PubMed).

Background publications Wilson, Disse-Nicodème, Choate et al.: "Human hypertension caused by mutations in WNK kinases." in: Science (New York, N.Y.), Vol. 293, Issue 5532, pp. 1107-12, 2001 (PubMed).

Catalog No. ABIN153167

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