CKAP4/p63 Antibody [Unconjugated]

Western blot shows lysates of JEG-3 human epithelial choriocarcinoma cell line, TK-6 human lymphoblast cell line, and HEK293 human embryonic kidney cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human CKAP4/p63 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7355) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for CKAP4/p63 at approximately 63 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

CKAP4/p63 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human CKAP4/p63 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7355) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

CKAP4/p63 was detected in immersion fixed NHEK human normal epidermal keratinocytes using Sheep Anti-Human CKAP4/p63 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7355) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 493-conjugated Anti-Sheep IgG Secondary Antibody (green; NL012). Cytokeratin 14 was also detected using Mouse Anti-Human Cytokeratin 14 Monoclonal Antibody (MAB3164) and stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007). Cells were counterstained with DAPI (blue). Specific staining of CKAP4/p63 was localized to perinuclear space and cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/ml. A specific band was detected for CKAP4/p63 at approximately 65 kDa (as indicated) using 4 µg/mL of Sheep Anti-Human CKAP4/p63 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7355). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

CKAP4/p63 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human CKAP4/p63
  His128-Val602
  Accession # Q07065
• Specificity: Detects human CKAP4/p63 in direct ELISAs and Western blots. In direct ELISAs, approximately 45% cross-reactivity with recombinant mouse CKAP4/p63 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: CKAP4
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Simple Western 4 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CKAP4/p63 Antibody [Unconjugated]

• 63 kDa membrane protein
• CKAP4
• CLIMP-63
• cytoskeleton-associated protein 4
• ERGIC-63
• MGC99554
• p63
• transmembrane protein (63kD), endoplasmic reticulum/Golgi intermediatecompartment
• type-II transmembrane protein p63

Background

CKAP4 (Cytoskeleton-associated protein 4; also CLIMP63 and p63) is a 63-64 kDa molecule that belongs to no known protein family. It is found intracellularly, and on the plasma membrane of select cells such as vascular smooth muscle and Type II Greater alveolar lung cells. CKAP4 has a bimodal distribution. First, it is embedded in the membrane of a compartment that links the ER with the Golgi apparatus. This localization is dependent upon its ability to form homooligomers, and its presence serves to anchor microtubules and direct the formation of tubular ER. Second, it is embedded in the plasma membrane and serves as a receptor for SP-A/surfactant protein-A (in lung) and tPA (in vessels). Human CKAP4 is a 602 amino acid (aa) type II transmembrane nonglycosylated protein. It contains a 106 aa N-terminal cytoplasmic region plus a 475 aa C-terminal luminal domain (aa 128-602). The luminal domain contains three coiled-coil regions (aa 130-214; 256-460; 533-602) plus three utilized phosphorylation sites. CKAP4 undergoes reversible palmitoylation. There are two potential isoform variants. One contains an alternative start at Met269, while another shows a deletion of aa 258-435. Over aa 128-602, human CKAP4 shares 82% aa sequence identity with mouse CKAP4.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CKAP4/p63 Antibody (AF7355)(3)

Publications using AF7355

• P Franzka, SC Schüler, T Kentache, R Storm, A Bock, I Katona, J Weis, K Buder, C Kaether, CA Hübner Impact of Hypermannosylation on the Structure and Functionality of the ER and the Golgi Complex Biomedicines, 2023-01-06;11(1):. 2023-01-06 [PMID: 36672654] (ICC, Mouse)
• Bowen Wang, Zhiheng Zhao, Michael Xiong, Rui Yan, Ke Xu The endoplasmic reticulum adopts two distinct tubule forms Proceedings of the National Academy of Sciences 5/3/2022 [PMID: 35471903]
• Bowen Wang, Zhiheng Zhao, Michael Xiong, Rui Yan, Ke Xu The endoplasmic reticulum adopts two distinct tubule forms Proceedings of the National Academy of Sciences 2022-05-03 [PMID: 35471903] (Immunocytochemistry, Primate - Chlorocebus aethiops (African Green Monkey))

Bioinformatics

• Gene Symbol: CKAP4
• Uniprot:
  • Q07065(Human)