CrkL Rabbit Polyclonal Antibody
cat.: HA500536
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC, IHC-P
Clonality: Polyclonal
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1.053ug/ul
Purification: Immunogen affinity purified.
Molecular weight: Predicted band size: 34 kDa
Isotype: IgG
Immunogen: Recombinant protein within human CrkL aa 1-303 / 303.
Positive control: K-562 cell lysate, 293T cell lysate, Jurkat cell lysate, HT-29 cell lysate, THP-1 cell lysate, A431 cell lysate, HL-60 cell lysate, C6 cell lysate, Mouse thymus tissue lysate, Rat thymus tissue lysate, K-562, human breast cancer tissue, human ovary cancer tissue, mouse cerebellum tissue, rat cerebellum tissue.
Subcellular location: Cytosol, extrinsic component of postsynaptic membrane, neuromuscular junction, nucleoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IHC-P
1:2,000
1:100
1:1,000
1:1,000
Uniprot #: SwissProt: P46109 Human | P47941 Mouse | Q5U2U2 Rat
Alternative names: Crk L Crk like protein Crk-like protein Crkl CRKL_HUMAN Crkol HGNC:2363 Oncogene CrkL V crk avian sarcoma virus CT10 oncogene homolog like v crk sarcoma virus CT10 oncogene homolog (avian) like V crk sarcoma virus CT10 oncogene homolog avian like
Images
HA500536_1.jpg Fig1: Western blot analysis of CrkL on different lysates with Rabbit anti-CrkL antibody (HA500536) at 1/2,000 dilution.

Lane 1: K-562 cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: HT-29 cell lysate (20 µg/Lane)
Lane 5: THP-1 cell lysate (20 µg/Lane)
Lane 6: A431 cell lysate (20 µg/Lane)
Lane 7: HL-60 cell lysate (low expression) (20 µg/Lane)
Lane 8: C6 cell lysate (20 µg/Lane)
Lane 9: Mouse thymus tissue lysate (40 µg/Lane)
Lane 10: Rat thymus tissue lysate (40 µg/Lane)

Predicted band size: 34 kDa
Observed band size: 37 kDa

Exposure time: 40 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 (HA500536) at 1/2,000 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.
HA500536_2.jpg Fig2: Immunocytochemistry analysis of K-562 cells labeling CrkL with Rabbit anti-CrkL antibody (HA500536) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-CrkL antibody (HA500536) 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 (M1305-2, 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.
HA500536_3.jpg Fig3: Flow cytometric analysis of K-562 cells labeling CrkL.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA500536, 1μg/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).
HA500536_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-CrkL antibody (HA500536) 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 (HA500536) 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.
HA500536_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue with Rabbit anti-CrkL antibody (HA500536) 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 (HA500536) 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.
HA500536_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-CrkL antibody (HA500536) 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 (HA500536) 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.
HA500536_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-CrkL antibody (HA500536) 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 (HA500536) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.