HA751426

CHD8 Recombinant Rabbit Monoclonal Antibody [PSH11-71]

cat.: HA751426

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, IP, ChIP
Clonality:	Monoclonal
Clone number:	PSH11-71

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 291 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human CHD8 aa 321-570.
Positive control:	HeLa cell lysate, SH-SY5Y cell lysate, 293T cell lysate, NIH/3T3 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa, NIH/3T3, C6, mouse testis tissue, rat testis tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P IP ChIP	1:1,000 1:50-1:100 1:100-1:200 1-2μg/sample Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #:	SwissProt: Q9HCK8 Human \| Q09XV5 Mouse \| Q9JIX5 Rat
Alternative names:	ATP dependent helicase CHD8 ATP-dependent helicase CHD8 Axis duplication inhibitor Beta catenin binding protein like CHD 8 CHD-8 chd8 CHD8_HUMAN Chromodomain helicase DNA binding protein 8 Chromodomain-helicase-DNA-binding protein 8 DKFZp686N17164 Duplin Helicase with SNF2 domain 1 HELSNF1 KIAA1564

Images

Fig1: Western blot analysis of CHD8 on different lysates with Rabbit anti-CHD8 antibody (HA751426) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: SH-SY5Y cell lysate (no heat)
Lane 3: 293T cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: Mouse brain tissue lysate
Lane 6: Rat brain tissue lysate
Lane 7: Rat brain tissue lysate (no heat)

Notice: no heat means the lysate is not boiled.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 291 kDa
Observed band size: 291 kDa

Exposure time: Lane 1-3: 14 seconds; Lane 4-7: 46 seconds; ECL: K1801;
3-8% 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 (HA751426) 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.

Fig2: Immunocytochemistry analysis of HeLa cells labeling CHD8 with Rabbit anti-CHD8 antibody (HA751426) 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-CHD8 antibody (HA751426) 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.

Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling CHD8 with Rabbit anti-CHD8 antibody (HA751426) at 1/50 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-CHD8 antibody (HA751426) at 1/50 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.

Fig4: Immunocytochemistry analysis of C6 cells labeling CHD8 with Rabbit anti-CHD8 antibody (HA751426) at 1/50 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-CHD8 antibody (HA751426) at 1/50 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.

	Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-CHD8 antibody (HA751426) 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 (HA751426) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-CHD8 antibody (HA751426) at 1/100 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 (HA751426) at 1/100 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.
	Fig7: CHD8 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA751426 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751426 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: HA751426 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA751426 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 59 seconds; ECL: K1801
	Fig8: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with CHD8 (HA751426) or 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”.