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Product Info Summary
| SKU: | PB9940 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IHC, IHC-F, ICC, WB |
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Product info
Product Name
Anti-Alpha A Crystallin/CRYAA Antibody Picoband®
SKU/Catalog Number
PB9940
PB0962 is an alternative SKU for this antibody, used in previous lots.
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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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 antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
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.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9940 is reactive to CRYAA in Human, Mouse, Rat
Observed Molecular Weight
20 kDa
Calculated molecular weight
19.9 kDa
Background of 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.
Application & Images
Applications
PB9940 is guaranteed for Flow Cytometry, IHC, IHC-F, ICC, WB Boster Guarantee
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
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Human
Immunocytochemistry, 0.5-1μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: rat spleen tissue, mouse eye ball tissue, SMMC-7721 whole cell
IHC: mouse spleen tissue, rat spleen tissue
FCM: HepG2 cell
Validation Images & Assay Conditions
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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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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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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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Flow Cytometry analysis of HepG2 cells using anti-CRYAA antibody (PB9940).
Overlay histogram showing HepG2 cells stained with PB9940 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-Alpha A Crystallin/CRYAA Antibody Picoband® (PB9940)
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1 Customer Q&As for Anti-Alpha A Crystallin/CRYAA Antibody Picoband®
Question
We are currently using anti-Alpha A Crystallin/CRYAA antibody PB9940 for rat tissue, and we are satisfied with the IHC-F results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on goat tissues as well?
Verified Customer
Verified customer
Asked: 2019-12-25
Answer
The anti-Alpha A Crystallin/CRYAA antibody (PB9940) has not been validated for cross reactivity specifically with goat tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in goat you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-12-25
