AlphaB Crystallin/CRYAB Antibody (731502)

Detection of Human, Mouse, and Rat AlphaB Crystallin/CRYAB by Western Blot.

Western blot shows lysates of rat heart tissue, mouse heart tissue, and human heart tissue. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat AlphaB Crystallin/CRYAB Monoclonal Antibody (Catalog # MAB4849) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). For additional reference, recombinant human AlphaB Crystallin/CRYAB and recombinant human AlphaA Crystallin/CRYAA (5 ng/lane) were included. A specific band was detected for AlphaB Crystallin/CRYAB at approximately 23 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human AlphaB Crystallin/CRYAB by Simple Western^TM.

Simple Western lane view shows lysates of human heart tissue, loaded at 0.2 mg/mL. A specific band was detected for AlphaB Crystallin/CRYAB at approximately 28 kDa (as indicated) using 1 µg/mL of Mouse Anti-Human/Mouse/Rat AlphaB Crystallin/CRYAB Monoclonal Antibody (Catalog # MAB4849). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of AlphaB Crystallin/CRYAB by Western Blot

BAG3 is recruited to mitochondria upon heat shock but is not imported into mitochondria.a, HeLa cells were subjected to heat shock (42 °C) for 1 h. Cells were collected afterward and the mitochondrial fraction was separated from the cytosolic fraction. Using immunoblotting we verified the abundance of BAG3 and sHSPs in each fraction. Tubulin was used as a marker for the cytosol and VDAC1 was used as a marker for mitochondria. The NP-40-soluble whole cell lysate (WCL) was used as a reference. b, Proteinase K protection assay on mitochondria isolated from HeLa cells. Isolated mitochondria were resuspended in a hypotonic (osmotic swelling) buffer to generate mitoplasts. Proteinase K (10 μg/ml) was added to digest accessible proteins. Samples were analyzed by SDS-PAGE followed by immunoblotting using anti-BAG3, anti-TOM20 (OM), anti-AIFM1 (IMS), anti-HCCS (IM), and anti-SDHA (matrix) antibodies. c, BAG3 knockout HeLa cells were rescued with wild type or delta-IPV BAG3-GFP (deficient in binding to HSPB8) and subjected to proteinase K protection assay as in panel B. Samples were analyzed by SDS-PAGE followed by immunoblotting using anti-BAG3, anti-TOM20 (OM), anti-AIFM1 (IMS), anti-HCCS (IM), and anti-HSP60 (matrix) antibodies. Results are representative of two (a,b) or three (c) replicates. Unprocessed blots are available in source data.Source data Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36690850), licensed under a CC-BY license. Not internally tested by R&D Systems.