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: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
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 (Human cervical adenocarcinoma cell) cell lysate, HEK-293 (Human embryonic kidney cell) cell lysate, MCF7 (Human breast cancer cell) cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, C6 (Rat glioma cell) cell lysate, PC-12, human prostate carcinoma tissue.
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
1:200-1:1,000
1:1,000
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 1/2,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: HEK-293 (Human embryonic kidney cell) cell lysate
Lane 3: MCF7 (Human breast cancer cell) cell lysate
Lane 4: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 5: C6 (Rat glioma cell) cell lysate

Lysates/proteins at 15 µg/Lane.
Exposure time: 1 minute 50 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ET1609-10, 1/2,000 in 5% NFDM/TBST, overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 36 kDa
Observed band size: 50 kDa
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 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_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Breast cancer
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1609-10, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1609-10_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Stomach
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1609-10, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1609-10_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Stomach
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1609-10, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1609-10_7.jpg Fig7: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Stomach
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1609-10, 1/1,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
ET1609-10_8.jpg Fig8: Application: Flow Cytometry (Intra)

Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: ET1609-10 (1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.