CLDN9 antibody - C-terminal region (ARP33617_T100)

Catalog#: ARP33617_T100

• Size: 100ug

Human HepG2

• Description of Target: CLDN9 is a member of claudin family. It is a four-transmembrane protein with WWCC motif, defined by W-X(17-22)-W-X(2)-C-X(8-10)-C.
• Gene Symbol: CLDN9
• Official Gene Full Name: Claudin 9
• Protein Accession#: NP_066192
• Nucleotide Accession#: NM_020982
• Swissprot Id: O95484
• Host: Rabbit
• Clonality: Polyclonal
• Protein Size (# AA): 217
• Molecular Weight: 24kDa
• Application: WB
• Immunogen: The immunogen for anti-CLDN9 antibody: synthetic peptide directed towards the C terminal of human CLDN9
• Product Format: Lyophilized powder
• Purification: Protein A purified
• Predicted Homology Based on Immunogen Sequence: Bovine: 100%; Dog: 100%; Mouse: 100%; Pig: 100%; Rat: 100%
• Peptide Sequence: Synthetic peptide located within the following region: WAAAALLMLGGGLLCCTCPPPQVERPRGPRLGYSIPSRSGASGLDKRDYV
• Blocking Peptide: For anti-CLDN9 antibody is Catalog # AAP33617 (Previous Catalog # AAPP04673)
• Key Reference: Katoh,M. and Katoh,M., (2003) Int. J. Mol. Med. 11 (6), 683-689
• Reconstitution and Storage: Add 100 μl of distilled water. Final anti-CLDN9 antibody concentration is 1 mg/ml in PBS buffer. For longer periods of storage, store at -20°C. Avoid repeat freeze-thaw cycles.

Product Protocols: SDS-PAGE and immunoblotting Protocol with CLDN9 Antibody (Catalog Number: ARP33617_T100)

Product Protocols:
SDS-PAGE and immunoblotting Protocol with Gene Name: CLDN9 Antibody (Catalog Number: ARP33617_T100)

Catalog Number: ARP33617_T100

Citation:
1. Abuazza G, Becker A, Williams SS, Chakravarty S, Truong HT, Lin F, Baum M. Claudins 6, 9, and 13 are developmentally expressed renal tight junction proteins. Am J Physiol Renal Physiol. 2006 Dec,291(6):F1132-41. Epub 2006 Jun 13.
PubMed PMID: 16774906.
Product Name: CLDN9 antibody - C-terminal region (ARP33617_T100)

Gene Name: CLDN9

Species: Mouse

Experiment Name: SDS-PAGE and immunoblotting

Experiment Background:
1. To determine whether there are developmental claudin isoforms, Ghazala et al. compared the claudin isoforms present in the neonatal and adult kidney using RT-PCR to detect mRNA of claudin isoforms. Claudin 6, claudin 9, and claudin 13 were either not expressed or barely detectable in the adult mouse kidney using traditional PCR, but were expressed in the neonatal mouse kidney.
2. Using real-time RT-PCR, we were able to detect a low level of claudin 6 mRNA expression in the adult kidney compared with the neonate, but claudin 9 and claudin 13 were only detected in the neonatalkidney. There was the same maturational decrease in these claudin proteins with Western blot analysis.
3. Claudin 9 was only consistently detectable in proximal convoluted tubules of 1-day-old neonates and only after more than 35 cycles of amplification. Claudin 13 was not detectable. It is possible that claudin 9 and 13 play a role in determining neonatal paracellular permeability in a distal nephron segment.

Experimental Steps:
1. Neonatal and adult kidney protein (100 ug/lane) were denatured and separated on a 6% polyacrylamide gel using SDS-PAGE.
2. The proteins were transferred overnight to a polyvinylidene diflouride membrane at 120-140 mA at 4 C.
3. The blot was blocked with fresh Blotto (5% nonfat milk and 0.1% Tween 20 in PBS, pH 7.4) for 1 h followed by incubation with overnight at 4 C with a primary antibody to mouse claudin 6 (Orbigen, San Diego, CA) at a 1:5,000 dilution, claudin 9 (Aviva Systems Biology, San Diego, CA) at 1:1,000 dilution or claudin 13 (Zymed Laboratories, South San Francisco, CA) at 1:100 dilution.
4. The blot was then washed extensively with Blotto.
5. The secondary antibody, horseradish peroxidase-conjugated donkey anti-rabbit immunoglobulin, was added at 1:10,000 dilution and incubated at room temperature for 1 h.
6. The blot was again washed with Blotto, and enhanced chemiluminescence was used to detect boundantibody (Amersham Biosciences, Piscataway, NJ). Claudin 6, 9, and 13 protein abundance was quantitated using densitometry.
7. Equal loading of samples was confirmed using an antibody to -actin at 1:5,000 dilution (Sigma Biochemicals, St. Louis, MO). There were at least six samples in each group.

Product Protocols: CLDN9 antibody tested with Human Hepg2 Cells (ARP33617_T100)

Aviva Systems Biology is the original manufacturer of this CLDN9 antibody (ARP33617_T100)

Click here to view the CLDN9 antibody Western Blot Protocol

Product Datasheet Link: CLDN9 antibody (ARP33617_T100)

WB Suggested Anti-CLDN9 Antibody Titration: 2.0ug/ml
Positive Control: HepG2

Western Blot image:


Description of Target: CLDN9 is a member of claudin family. It is a four-transmembrane protein with WWCC motif, defined by W-X(17-22)-W-X(2)-C-X(8-10)-C.

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Aviva Systems Biology’s CLDN9 antibody (ARP33617_T100) has been tested using other cell lysates and tissues. To obtain more data about this antibody please email us at info@avivasysbio.com.

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