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82941
AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)
Antibody Conjugates
Monoclonal Antibody

AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #82941

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Immunofluorescence Image 1: AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)

Confocal immunofluorescent analysis of mouse cerebellum using AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) (green) and Myelin Basic Protein (D8X4Q) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #30103 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

Immunofluorescence Image 2: AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)

Confocal immunofluorescent analysis of mouse colon using AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

Immunofluorescence Image 3: AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)

Confocal immunofluorescent analysis of mouse kidney using AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).

To Purchase # 82941S
Product # Size Price Inventory
82941S
100 µl  (50 tests)

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Supporting Data

REACTIVITY H M R
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Rabbit IgG

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct immunofluorescent analysis in mouse tissues. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated AQP4 (D1F8E) XP® Rabbit mAb #59678.

Product Usage Information

Application Dilution
Immunofluorescence (Frozen) 1:50

Storage

Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Immunofluorescence (Frozen)

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (9808) To prepare 1L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix. Adjust pH to 8.0.
  2. Formaldehyde: 16%, methanol free, Polysciences, Inc. (cat# 18814), use fresh and store opened vials at 4°C in dark, dilute in 1X PBS for use.
  3. Blocking Buffer (1X PBS / 5% normal goat serum (#5425) / 0.3% Triton™ X-100): To prepare 10 ml: add 0.5 ml normal goat serum and 0.5 ml 20X PBS to 9.0 ml dH2O, mix. While stirring, add 30 µl Triton™ X-100.
  4. Antibody Dilution Buffer: (1X PBS / 1% BSA / 0.3% Triton™ X-100): To prepare 10 ml, add 30 µl Triton™ X-100 to 10 ml 1X PBS. Mix well then add 0.1 g BSA (#9998), mix.
  5. Prolong® Gold AntiFade Reagent (#9071), Prolong® Gold AntiFade Reagent with DAPI (#8961).

B. Specimen Preparation - Frozen/Cryostat Sections (IF-F)

  1. For fixed frozen tissue proceed with Immunostaining (Section C).
  2. For fresh, unfixed frozen tissue, please fix immediately, as follows:
    1. Cover sections with 4% formaldehyde dilute in 1X PBS.

      NOTE: Formaldehyde is toxic, use only in fume hood.

    2. Allow sections to fix for 15 minutes at room temperature.
    3. Aspirate liquid, rinse three times in 1X PBS for 5 minutes each.
    4. Proceed with Immunostaining (Section C).

C. Immunostaining

NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.

  1. Block specimen in Blocking Buffer for 60 min.
  2. While blocking, prepare primary antibody by diluting as indicated on protocol on product webpage in Antibody Dilution Buffer.
  3. Aspirate blocking solution, apply diluted primary antibody.
  4. Incubate overnight at 4°C.
  5. Rinse three times in 1X PBS for 5 min each.
  6. Coverslip slides with Prolong® Gold Antifade Reagent (#9071) or Prolong® Gold Antifade Reagent with DAPI (#8961).
  7. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 4°C protected from light.

posted November 2006

revised November 2013

Protocol Id: 222

Specificity / Sensitivity

AQP4 (D1F8E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) recognizes endogenous levels of total AQP4 protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the carboxy terminus of human AQP4 protein.

Background

Aquaporins (AQP) are integral membrane proteins that serve as channels in the transfer of water and small solutes across the membrane. There are 13 isoforms of AQP that express in different types of cells and tissues (1,2). AQP1 is found in blood vessels, kidney, eye, and ear. AQP2 is found in the kidney, and it has been shown that the lack of AQP2 results in diabetes (1,3). AQP4 is present in the brain, where it is enriched in astrocytes (1,2,4). AQP5 is found in the salivary and lacrimal gland, AQP6 in intracellular vesicles in the kidney, AQP7 in adipocytes, AQP8 in kidney, testis, and liver, AQP9 is present in liver and leukocytes and AQP10-11 in the intestine (1,3,4). AQPs are essential for the function of cells and organs. It has been shown that AQP1 and AQP4 regulate the water homeostasis in astrocytes, preventing cerebral edema caused by solute imbalance (5). Several studies have shown the involvement of AQPs in the development of inflammatory processes, including cells of innate and adaptive immunity (6,7).

  1. Takata, K. et al. (2004) Prog Histochem Cytochem 39, 1-83.
  2. Echevarría, M. and Ilundáin, A.A. (1998) J Physiol Biochem 54, 107-18.
  3. Gomes, A. et al. (2018) Front Chem 6, 238.
  4. Xu, M. et al. (2017) Adv Exp Med Biol 969, 81-103.
  5. Kobayashi, H. et al. (2004) J Pharmacol Sci 96, 264-70.
  6. Meli, R. et al. (2018) Front Physiol 9, 101.
  7. Ishibashi, K. et al. (1998) Biochem Biophys Res Commun 244, 268-74.

Pathways & Proteins

Explore pathways + proteins related to this product.

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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
ProLong is a registered trademark of Life Technologies Corporation.
This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected]