PHB2 Rabbit Polyclonal Antibody

SKU: orb185934

Description

PHB2 Rabbit Polyclonal Antibody

Images & Validation

• Tested Applications: FC, ICC, WB
• Dilution range: WB=1:500-2000, ICC/IF=1:100-500, Flow-Cyt=2ug/Test
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Bovine, Equine, Sheep

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human PHB2 (281-299/299aa)
• Target: PHB2
• Molecular Weight: 33 kDa
• Purification: Affinity purified by Protein A
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Form/Appearance: Liquid
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Concentration: 1mg/ml
• Disclaimer: For research use only

Alternative Names

PHB2_HUMAN; Prohibitin-2; B-cell receptor-associated protein BAP37; D-prohibitin; Repressor of estrogen receptor activity; BAP; REA; p22; hBAP; Bap37; BCAP37; PNAS-141;

Blank control (black line): Hela. Primary Antibody (green line): Rabbit Anti-PHB2 antibody (orb185934), dilution: 2 ug/Test, Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/Test. Isotype control (orange line): Normal Rabbit IgG, Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Hela cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (PHB2) polyclonal Antibody, Unconjugated (orb185934) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Protein: spleen (mouse) lysate at 40 ug, Primary: rabbit Anti-REA (orb185934) at 1:300, Secondary: HRP conjugated Goat-Anti-rabbit IgG (orb572747) at 1:5000, Predicted band size: 33 kD, Observed band size: 35 kD.

Sample: Lane 1: Mouse Kidney tissue lysates, Lane 2: Mouse Testis tissue lysates, Lane 3: Mouse NIH/3T3 cell lysates, Lane 4: Rat Kidney tissue lysates, Lane 5: Rat Testis tissue lysates, Lane 6: Human HeLa cell lysates, Lane 7: Human 293T cell lysates, Lane 8: Human HT-1080 cell lysates, Lane 9: Human A431 cell lysates, Lane 10: Human A549 cell lysates, Primary: Anti-PHB2 (orb185934) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 33 kDa, Observed band size: 35 kDa.

Tissue/Cell: colonic epithelial cells, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-REA Polyclonal Antibody, Unconjugated (orb185934) 1:100, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Quick Database Links

Gene Symbol

PHB2

UniProt

Q99623