Eph receptor B2 Recombinant Rabbit Monoclonal Antibody [PSH23-74]
cat.: HA724397
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-Fr, IP
Clonality: Monoclonal
Clone number: PSH23-74
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 117 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Eph receptor B2 aa 1-543.
Positive control: COLO205 (Human colon adenocarcinoma cells) cell lysate, K-562 (Human chronic myelogenous leukemia cells) cell lysate, Saos-2 (Human osteosarcoma cells) cell lysate, NIH/3T3 (Mouse fibroblasts) cell lysate, 4T1 (Mouse breast cancer cells) cell lysate, Mouse brain tissue lysate, Mouse colon tissue lysate, Rat brain tissue lysate, Rat colon tissue lysate.
Subcellular location: Cell membrane, Cell projection, axon, dendrite.
Recommended Dilutions:
  WB
  IHC-Fr
  IP

1:5,000
1:500
1-2μg/sample
Uniprot #: SwissProt: P29323 Human | P54763 Mouse
Entrez Gene: 313633 Rat
Alternative names: cek5 Developmentally regulated EPH related tyrosine kinase DRT EK5 ELK related protein tyrosine kinase Eph receptor B2 EPH tyrosine kinase 3 EPH-like kinase 5 EPHB2 EPHB2_HUMAN Ephrin type B receptor 2 Ephrin type-B receptor 2 EPHT 3 ERK ETECK hEK5 Nuk Prkm 5 Receptor protein tyrosine kinase HEK 5 Renal carcinoma antigen NY-REN-47 Sek 3 Tyro 5 Tyrosine protein kinase receptor CEK 5 Tyrosine protein kinase receptor EPH 3 Tyrosine protein kinase receptor QEK 5 Tyrosine-protein kinase receptor EPH-3 Tyrosine-protein kinase TYRO5
Images
HA724397_1.jpg Fig1: Western blot analysis of Eph receptor B2 on different lysates with Rabbit anti-Eph receptor B2 antibody (HA724397) at 1/5,000 dilution.

Lane 1: COLO205 (Human colon adenocarcinoma cells) cell lysate
Lane 2: K-562 (Human chronic myelogenous leukemia cells) cell lysate
Lane 3: Saos-2 (Human osteosarcoma cells) cell lysate
Lane 4: NIH/3T3 (Mouse fibroblasts) cell lysate
Lane 5: 4T1 (Mouse breast cancer cells) cell lysate
Lane 6: Mouse brain tissue lysate
Lane 7: Mouse colon tissue lysate
Lane 8: Rat brain tissue lysate
Lane 9: Rat colon tissue lysate

Cell lysates/proteins at 20 µg/Lane. Tissue lysates/proteins at 40 µg/Lane.
Exposure time: 3 minutes; ECL: K1801

K-562 is a negative control (PMID : 18713744, PMID : 21508957).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724397, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 117 kDa
Observed band size: 130 kDa
HA724397_2.jpg Fig2: Application: Immunofluorescence (IHC-Fr)

Species: Mouse
Tissue: Colon
Sample: Frozen section

Antigen retrieval: Not required

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 0.5 % Triton X-100 + 0.3 M Glycine in PBS, 10 minutes at room temperature.
Primary antibody: HA724397, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA724397_3.jpg Fig3: Application: Immunofluorescence (IHC-Fr)

Species: Rat
Tissue: Colon
Sample: Frozen section

Antigen retrieval: Not required

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 0.5 % Triton X-100 + 0.3 M Glycine in PBS, 10 minutes at room temperature.
Primary antibody: HA724397, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA724397_4.jpg Fig4: Immunoprecipitation (IP)

Eph receptor B2 was immunoprecipitated in 0.2 mg COLO205 (Human colon adenocarcinoma cells) cell lysate with HA724397 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724397 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: COLO205 cell lysate (input)
Lane 2: HA724397 IP in COLO205 cell lysate
Lane 3: Rabbit IgG instead of HA724397 in COLO205 cell lysate

Exposure time: 2 minutes 6 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA724397, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 117 kDa
Observed band size: 130 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.