YB1 Recombinant Rabbit Monoclonal Antibody [ST0432]
cat.: ET1609-10
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: ST0432
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 36 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human YB1 aa 275-324 / 324.
Positive control: HeLa cell lysate, Jurkat cell lysate, PC-12, human kidney tissue, mouse colon tissue, mouse stomach tissue, human prostate carcinoma tissue, Hela.
Subcellular location: Cytoplasm, Nucleus, Cytoplasmic granule, Secreted.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:5,000
1:100-1:500
1:100-1:500
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P67809 Human | P62960 Mouse | P62961 Rat
Alternative names: BP 8 CBF-A CCAAT binding transcription factor I subunit A CCAAT-binding transcription factor I subunit A CSDA2 CSDB DBPB DNA binding protein B DNA-binding protein B EFI-A Enhancer factor I subunit A MDR NF1 MGC104858 MGC110976 MGC117250 NSEP 1 NSEP1 Nuclease sensitive element binding protein 1 Nuclease-sensitive element-binding protein 1 p50 Q15905 Y-box binding protein 1 Y-box transcription factor Y-box-binding protein 1 YB 1 YB-1 YBOX1_HUMAN YBX 1 ybx1
Images
ET1609-10_1.jpg Fig1: Western blot analysis of YB1 on different lysates with Rabbit anti-YB1 antibody (ET1609-10) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 36 kDa
Observed band size: 50 kDa

Exposure time: 2 minutes 18 seconds;
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 (ET1609-10) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1609-10_2.jpg Fig2: Immunocytochemistry analysis of PC-12 cells labeling YB1 with Rabbit anti-YB1 antibody (ET1609-10) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-YB1 antibody (ET1609-10) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1609-10_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-YB1 antibody at 1/200. Counter stained with hematoxylin.
ET1609-10_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-YB1 antibody at 1/200. Counter stained with hematoxylin.
ET1609-10_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue using anti-YB1 antibody at 1/200. Counter stained with hematoxylin.
ET1609-10_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue with Rabbit anti-YB1 antibody (ET1609-10) 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 (ET1609-10) 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.
ET1609-10_7.jpg Fig7: Flow cytometric analysis of Hela cells with YB1 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.