ADNP Recombinant Rabbit Monoclonal Antibody [PSH24-23]
cat.: HA751933
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-Fr, IHC-P, IF-Cell, IP, ChIP
Clonality: Monoclonal
Clone number: PSH24-23
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 124 kDa
Isotype: IgG
Immunogen: Recombinant protein within human ADNP aa 803-1,102.
Positive control: HeLa (Human cervical adenocarcinoma cell) cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate, C2C12 (Mouse myoblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, C6 (Rat glioma cell) cell lysate.
Subcellular location: Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IHC-Fr
  IHC-P
  IF-Cell
  IP
  ChIP
1:2,000
1:500
1:2,000
1:100-1:500
1-2μg/sample
Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #: SwissProt: Q9H2P0 Human | Q9Z103 Mouse | Q9JKL8 Rat
Alternative names: Activity dependent neuroprotective protein Activity dependent neuroprotector Activity-dependent neuroprotective protein Activity-dependent neuroprotector homeobox protein Adnp ADNP_HUMAN KIAA0784 ADNP1
Images
HA751933_1.jpg Fig1: Western blot analysis of ADNP on different lysates with Rabbit anti-ADNP antibody (HA751933) at 1/5,000 dilution.

Lane 1: 293T-ADNP-WT cell lysate
Lane 2: 293T-ADNP-KO cell lysate
Lane 3: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 4: NIH/3T3 (Mouse fibroblast) cell lysate
Lane 5: C2C12 (Mouse myoblast) cell lysate
Lane 6: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
Lane 7: C6 (Rat glioma cell) cell lysate

Lysates/proteins at 20 µg/Lane.
Exposure time: 1 minute; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA751933, 1/2,000 in 5% NFDM/TBST, 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: 124 kDa
Observed band size: 140 kDa
HA751933_2.jpg Fig2: Application: Immunofluorescence (IHC-Fr)

Species: Mouse
Tissue: Brain
Sample: Frozen section

Antigen retrieval: Not required

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

Primary antibody: HA751933, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA751933_3.jpg Fig3: Application: Immunofluorescence (IHC-Fr)

Species: Rat
Tissue: Brain
Sample: Frozen section

Antigen retrieval: Not required

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

Primary antibody: HA751933, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA751933_4.jpg Fig4: Application: Immunohistochemistry (IHC-P)

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

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

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: HA751933, 1/2,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751933_5.jpg Fig5: Application: Immunohistochemistry (IHC-P)

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

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

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: HA751933, 1/2,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751933_6.jpg Fig6: Application: Immunohistochemistry (IHC-P)

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

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

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: HA751933, 1/2,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA751933_7.jpg Fig7: Application: Immunocytochemistry (IF-cell)

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

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751933, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA751933_8.jpg Fig8: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: Neuro-2a (Mouse brain neuroblastoma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751933, 1/100, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA751933_9.jpg Fig9: Application: Immunocytochemistry (IF-cell)

Species: Rat
Sample: C6 (Rat glioma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA751933, 1/500, overnight at 4°C.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA751933_10.JPG Fig10: Immunoprecipitation (IP)

ADNP was immunoprecipitated in 0.2 mg Neuro-2a (Mouse brain neuroblastoma cell) cell lysate with HA751933 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751933 at 1/1,000 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: Neuro-2a cell lysate (input)
Lane 2: HA751933 IP in Neuro-2a cell lysate
Lane 3: Rabbit IgG instead of HA751933 in Neuro-2a cell lysate

Exposure time: 59 seconds
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA751933, 1/1,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 124 kDa
Observed band size: 140 kDa
HA751933_11.jpg Fig11: Chromatin immunoprecipitations were performed with cross-linked chromatin from Neuro-2a cells with ADNP (HA751933) / Competitor's antibody / Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.