REA Recombinant Rabbit Monoclonal Antibody [JE35-46]
cat.: HA721588
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE35-46
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 33 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human REA aa 151-200 / 299.
Positive control: Huh7 cell lysate, HeLa cell lysate, HepG2 cell lysate, PANC-1 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, mouse heart tissue lysate, rat heart tissue lysate, mouse testis tissue lysate, rat testis tissue lysate, human liver tissue, rat liver tissue, HeLa.
Subcellular location: Mitochondrion inner membrane, Cytoplasm, Nucleus, Cell membrane.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  FC
1:1,000
1:200-1:1,000
1:100
1:500-1:1,000
Uniprot #: SwissProt: Q99623 Human | O35129 Mouse | Q5XIH7 Rat
Alternative names: B cell receptor associated protein BAP37 B-cell receptor-associated protein BAP37 BAP Bap37 BCAP 37 D prohibitin D-prohibitin p22 Phb2 PHB2_HUMAN PNAS 141 Prohibitin 2 Prohibitin-2 Repressor of estrogen receptor activity
Images
HA721588_1.jpg Fig1: Western blot analysis of REA on different lysates with Rabbit anti-REA antibody (HA721588) at 1/1,000 dilution.

Lane 1: Huh7 cell lysate (15 µg/Lane)
Lane 2: HeLa cell lysate (15 µg/Lane)
Lane 3: HepG2 cell lysate (15 µg/Lane)
Lane 4: PANC-1 cell lysate (15 µg/Lane)
Lane 5: RAW264.7 cell lysate (15 µg/Lane)
Lane 6: NIH/3T3 cell lysate (15 µg/Lane)
Lane 7: PC-12 cell lysate (15 µg/Lane)
Lane 8: Mouse liver tissue lysate (30 µg/Lane)
Lane 9: Rat liver tissue lysate (30 µg/Lane)
Lane 10: Mouse heart tissue lysate (30 µg/Lane)
Lane 11: Rat heart tissue lysate (30 µg/Lane)
Lane 12: Mouse testis tissue lysate (30 µg/Lane)
Lane 13: Rat testis tissue lysate (30 µg/Lane)

Predicted band size: 33 kDa
Observed band size: 33 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721588) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HA721588_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-REA antibody (HA721588) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721588) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721588_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-REA antibody (HA721588) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721588) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721588_4.jpg Fig4: Immunocytochemistry analysis of HeLa cells labeling REA with Rabbit anti-REA antibody (HA721588) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-REA antibody (HA721588) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA721588_5.jpg Fig5: Flow cytometric analysis of HeLa cells labeling REA.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA721588, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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