Anti-Alpha A Crystallin/CRYAA Antibody Picoband™

Product Name

Anti-Alpha A Crystallin/CRYAA Antibody Picoband™

View all AlphaA Crystallin/CRYAA Antibodies

SKU/Catalog Number

PB9940

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Alpha A Crystallin/CRYAA Antibody Picoband™ catalog # PB9940. Tested in Flow Cytometry, IHC, IHC-F, ICC, WB applications. This antibody reacts with Human, 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-Alpha A Crystallin/CRYAA Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9940)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E. coli-derived human Alpha A Crystallin recombinant protein (Position: M1-S173). Human Alpha A Crystallin shares 94.8% amino acid (aa) sequence identity with both mouse and rat Alpha A Crystallin.

*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

PB9940 is reactive to CRYAA in Human, Mouse, Rat

Applications

PB9940 is guaranteed for Flow Cytometry, IHC, IHC-F, ICC, WB Boster Guarantee

Observed Molecular Weight

20 kDa

Calculated molecular weight

19909 MW

Background of AlphaA Crystallin/CRYAA

Alpha-crystallin A chain is a protein that in humans is encoded by the CRYAA gene. Mammalian lens crystallins are divided into alpha, beta, and gamma families. Alpha crystallins are composed of two gene products: alpha-A and alpha-B, for acidic and basic, respectively. Alpha crystallins can be induced by heat shock and are members of the small heat shock protein (HSP20) family. They act as molecular chaperones although they do not renature proteins and release them in the fashion of a true chaperone; instead they hold them in large soluble aggregates. Two additional functions of alpha crystallins are an autokinase activity and participation in the intracellular architecture. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. Alpha-A and alpha-B gene products are differentially expressed; alpha-A is preferentially restricted to the lens and alpha-B is expressed widely in many tissues and organs. Defects in this gene cause autosomal dominant congenital cataract (ADCC).

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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Reconstitution

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, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat, Human, By Heat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CRYAA using anti-CRYAA antibody (PB9940).
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: rat spleen tissue lysates,
Lane 2: mouse eye ball tissue lysates,
Lane 3: SMMC-7721 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-CRYAA antigen affinity purified polyclonal antibody (Catalog # PB9940) 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 1:10000 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 CRYAA at approximately 20KD. The expected band size for CRYAA is at 20KD.

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Figure 2. IHC analysis of CRYAA using anti-CRYAA antibody (PB9940).
CRYAA was detected in paraffin-embedded section of mouse spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRYAA Antibody (PB9940) 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 CRYAA using anti-CRYAA antibody (PB9940).
CRYAA was detected in paraffin-embedded section of rat spleen tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CRYAA Antibody (PB9940) 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 HepG2 cells using anti-CRYAA antibody (PB9940).
Overlay histogram showing HepG2 cells stained with PB9940 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CRYAA Antibody (PB9940,1μg/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 CRYAA (Source: Uniprot.org, NCBI)

Alternative Names

AlphaA Crystallin; CRYA1; CRYA1crystallin, alpha-1; CRYAA; crystallin, alpha A; Heat shock protein beta-4; HSPB4; HSPB4alpha-crystallin A chain; human alphaA-crystallin (CRYA1), 10HspB4 CRYAA CRYA1, CTRCT9, HSPB4 crystallin alpha A alpha-crystallin A chain|crystallin, alpha-1|heat shock protein beta-4|human alphaA-crystallin (CRYA1)

For more info on CRYAA, check out the CRYAA Infographic

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In this infographic, you will see the following information for CRYAA: 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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