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Product Info Summary
| SKU: | A03496-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Alpha B Crystallin/CRYAB Antibody Picoband®
SKU/Catalog Number
A03496-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Alpha B Crystallin/CRYAB Antibody Picoband® catalog # A03496-2. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey, 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. CRYAB (alpha B-crystallin) is described as a small heat shock protein (HSP20 family) that can be heat-induced and acts as a molecular ‘holdase’ chaperone forming large soluble aggregates; additional functions noted include autokinase activity and participation in intracellular architecture. Assay context: antibody validated for ELISA, Flow Cytometry, IF, IHC, ICC, and WB; reactive to CRYAB in human/monkey/mouse/rat; immunogen is E. coli–derived human CRYAB recombinant protein (E30–Q151) with reported observed MW ~20 kDa and stated no cross-reactivity. Commonly interpreted in broader proteostasis/stress programs with autophagy regulators such as ATG5 and survival/cell-cycle markers such as BIRC5 (Survivin) (putative); tissue co-localization patterns can be explored in multiplex IHC workflows (putative).
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 B Crystallin/CRYAB Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03496-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Alpha B Crystallin/CRYAB recombinant protein (Position: E30-Q151).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03496-2 is reactive to CRYAB in Human, Monkey, Mouse, Rat
Observed Molecular Weight
20 kDa
Calculated molecular weight
20.2 kDa
Background of CRYAB
Alpha-crystallin B chain is a protein that in humans is encoded by the CRYAB 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. These heterogeneous aggregates consist of 30-40 subunits; the alpha-A and alpha-B subunits have a 3:1 ratio, respectively. 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. Elevated expression of alpha-B crystallin occurs in many neurological diseases; a missense mutation cosegregated in a family with a desmin-related myopathy. Alternative splicing results in multiple transcript variants.
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
A03496-2 is guaranteed for ELISA, Flow Cytometry, IF, IHC, 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, 0.1-0.25μg/ml, Mouse, Rat, Monkey
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: rat eye tissue, mouse eye tissue, rat heart tissue, mouse heart tissue, monkey heart tissue
IHC: rat brain tissue, human lung cancer tissue, human gastric cancer tissue, human skin cancer tissue, human glioma tissue, human gastric cancer tissue, mouse heart tissue, mouse brain tissue, mouse brain tissue, rat eye ball tissue
ICC/IF: U20S cell
FCM: THP-1 cell
Validation Images & Assay Conditions
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Western blot analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
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 eye tissue lysates,
Lane 2: mouse eye tissue lysates,
Lane 3: rat heart tissue lysates,
Lane 4: mouse heart tissue lysates,
Lane 5: monkey heart tissue 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-Alpha B Crystallin/CRYAB antigen affinity purified polyclonal antibody (Catalog # A03496-2) at 0.25 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 Alpha B Crystallin/CRYAB2 at approximately 20KD. The expected band size for Alpha B Crystallin/CRYAB is at 20KD.
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IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of rat brain 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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 Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of human lung cancer 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of human gastric cancer 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of human skin cancer 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of human glioma 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of human gastric cancer 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of mouse heart 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of mouse brain 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of mouse brain 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IHC analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in paraffin-embedded section of rat eye ball 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 2μg/ml rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) 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.
Click image to see more details
IF analysis of Alpha B Crystallin/CRYAB using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Alpha B Crystallin/CRYAB was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-Alpha B Crystallin/CRYAB Antibody (A03496-2) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of THP-1 cells using anti-Alpha B Crystallin/CRYAB antibody (A03496-2).
Overlay histogram showing THP-1 cells stained with A03496-2 (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-Alpha B Crystallin/CRYAB Antibody (A03496-2, 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 B Crystallin/CRYAB Antibody Picoband® (A03496-2)
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