Anti-Carbonic Anhydrase 4/CA4 Picoband™ Antibody

Product Name

Anti-Carbonic Anhydrase 4/CA4 Picoband™ Antibody

View all Carbonic Anhydrase IV/CA4 Antibodies

SKU/Catalog Number

A01523-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Carbonic Anhydrase 4/CA4 Picoband™ Antibody catalog # A01523-3. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-Carbonic Anhydrase 4/CA4 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01523-3)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse Carbonic Anhydrase 4/CA4 recombinant protein (Position: E25-L291).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A01523-3 is reactive to CA4 in Mouse, Rat

Applications

A01523-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

35 kDa

Calculated molecular weight

35.032kDa

Background of Carbonic Anhydrase IV/CA4

Carbonic anhydrase 4 is an enzyme that in humans is encoded by the CA4 gene. It is mapped to 11; 11 C. Carbonic anhydrases (CAs) are a large family of zinc metalloenzymes that catalyze the reversible hydration of carbon dioxide. They participate in a variety of biological processes, including respiration, calcification, acid-base balance, bone resorption, and the formation of aqueous humor, cerebrospinal fluid, saliva, and gastric acid. They show extensive diversity in tissue distribution and in their subcellular localization. This gene encodes a glycosylphosphatidyl-inositol-anchored membrane isozyme expressed on the luminal surfaces of pulmonary (and certain other) capillaries and proximal renal tubules. Its exact function is not known; however, it may have a role in inherited renal abnormalities of bicarbonate transport.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Mouse
Direct ELISA, 0.1-0.5μg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Ca4 using anti-Ca4 antibody (A01523-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: mouse lung tissue lysates,
Lane 2: rat PC-12 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Ca4 antigen affinity purified polyclonal antibody (Catalog # A01523-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Ca4 at approximately 35KD. The expected band size for Ca4 is at 35KD.

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Figure 2. IHC analysis of Ca4 using anti-Ca4 antibody (A01523-3).
Ca4 was detected in paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Ca4 Antibody (A01523-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of Ca4 using anti-Ca4 antibody (A01523-3).
Ca4 was detected in paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Ca4 Antibody (A01523-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of mouse spleen cells using anti-Ca4 antibody (A01523-3).
Overlay histogram showing mouse spleen cells stained with A01523-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Ca4 Antibody (A01523-3, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For CA4 (Source: Uniprot.org, NCBI)

Alternative Names

CA4; CAIV; CA-IV; Car4; Carbonate dehydratase IV; carbonic anhydrase 4; Carbonic Anhydrase IV; carbonic anhydrase IVRP17; carbonic dehydratase IV; EC 4.2.1.1; retinitis pigmentosa 17 (autosomal dominant); RP17 CA4 CAIV, Car4, RP17 carbonic anhydrase 4 carbonic anhydrase 4|CA-IV|carbonate dehydratase IV|carbonic anhydrase IV|carbonic dehydratase IV

For more info on CA4, check out the CA4 Infographic

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In this infographic, you will see the following information for CA4: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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