CHMP2B Recombinant Rabbit Monoclonal Antibody [JE54-35]
cat.: ET7110-67
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE54-35
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: 24 kDa.
Isotype: IgG
Immunogen: Recombinant protein within Human CHMP2B aa 103-213 / 213.
Positive control: Mouse bone marrow tissue lysate, rat bone marrow tissue lysate, human skeletal muscle tissue lysate, A549 cell lysate, A431 cell lysate, Hela cell lysate, SW620, SW1990, mouse testis tissue, mouse kidney tissue, A549.
Subcellular location: Late endosome membrane, cytosol.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:500-1:1,000
1:50-1:100
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: Q9UQN3 Human | Q8BJF9 Mouse
Alternative names: ALS17 Charged multivesicular body protein 2b CHM2B_HUMAN CHMP family, member 2B CHMP2.5 CHMP2b Chromatin modifying protein 2b Chromatin-modifying protein 2b hVps2-2 Vacuolar protein sorting 2, yeast, homolog of, B Vacuolar protein sorting 2-2 Vacuolar protein sorting-associated protein 2-2 VPS2 homolog B Vps2-2 VPS2B
Images
ET7110-67_1.jpg Fig1: Western blot analysis of CHMP2B on different lysates with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-CHMP2B KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 30 kDa

Exposure time: 30 seconds; ECL: K1802;

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 (ET7110-67) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET7110-67_2.jpg Fig2: Western blot analysis of CHMP2B on different lysates with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/500 dilution.

Lane 1: Mouse bone marrow tissue lysate
Lane 2: Rat bone marrow tissue lysate
Lane 3: Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 30 kDa

Exposure time: 2 minutes;

15% 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 (ET7110-67) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET7110-67_3.jpg Fig3: Western blot analysis of CHMP2B on different lysates with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/500 dilution.

Lane 1: A549 cell lysate
Lane 2: A431 cell lysate
Lane 3: Hela cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 30 kDa

Exposure time: 2 minutes;

15% 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 (ET7110-67) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET7110-67_4.jpg Fig4: Immunocytochemistry analysis of SW620 cells labeling CHMP2B with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/50 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-CHMP2B antibody (ET7110-67) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET7110-67_5.jpg Fig5: Immunocytochemistry analysis of SW1990 cells labeling CHMP2B with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/50 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-CHMP2B antibody (ET7110-67) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET7110-67_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET7110-67) at 1/100 dilution for 0.5 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.
ET7110-67_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-CHMP2B antibody (ET7110-67) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET7110-67) at 1/100 dilution for 0.5 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.
ET7110-67_8.jpg Fig8: Flow cytometric analysis of CHMP2B was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-67, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.