ET1706-22

DDB1 Recombinant Rabbit Monoclonal Antibody [JU32-35]

cat.: ET1706-22

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP
Clonality:	Monoclonal
Clone number:	JU32-35

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
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: 127 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human DDB1 aa 1040-1140 / 1,140.
Positive control:	PC-12 cell lysates, HepG2 cell lysate, NIH/3T3 cell lysate, MCF-7 cell lysate, rat kidney tissue lysate, Hela, HUVEC, SH-SY5Y, rat esophagus tissue, human liver carcinoma tissue, human kidney tissue, mouse brain tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IP	1:500-1:2,000 1:50-1:200 1:50-1:200 1:50-1:200 1:10-1:50
Uniprot #:	SwissProt: Q16531 Human \| Q3U1J4 Mouse \| Q9ESW0 Rat
Alternative names:	Damage specific DNA binding protein 1 Damage-specific DNA-binding protein 1 DDB 1 DDB p127 subunit Ddb1 DDB1_HUMAN DDBa DNA damage binding protein 1 DNA damage-binding protein 1 DNA damage-binding protein a HBV X-associated protein 1 UV damaged DNA binding factor UV damaged DNA binding protein 1 UV DDB 1 UV DDB1 UV-damaged DNA-binding factor UV-damaged DNA-binding protein 1 UV-DDB 1 X associated protein 1 XAP 1 XAP-1 XAP1 Xeroderma pigmentosum group E complementing protein Xeroderma pigmentosum group E-complementing protein XPCe XPE XPE BF XPE binding factor XPE-BF XPE-binding factor

Images

Fig1: Western blot analysis of DDB1 on PC-12 cell lysates with Rabbit anti-DDB1 antibody (ET1706-22) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 127 kDa
Observed band size: 127 kDa

Exposure time: 2 minutes;

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 (ET1706-22) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of DDB1 on different lysates with Rabbit anti-DDB1 antibody (ET1706-22) at 1/500 dilution.

Lane 1: HepG2 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: MCF-7 cell lysate
Lane 4: Rat kidney tissue lysate(20 µg/Lane)

Lysates/proteins at 10 µg/Lane.

Predicted band size: 127 kDa
Observed band size: 127 kDa

Exposure time: 2 minutes;

6% 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 (ET1706-22) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 200,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of Hela cells labeling DDB1 with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
	Fig4: Immunocytochemistry analysis of HUVEC cells labeling DDB1 with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
	Fig5: Immunocytochemistry analysis of SH-SY5Y cells labeling DDB1 with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution.Nuclear DNA was labelled in blue with DAPI.
	Fig6: Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 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 (ET1706-22) at 1/50 dilution for 0.5 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: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 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 (ET1706-22) at 1/50 dilution for 0.5 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 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 (ET1706-22) at 1/50 dilution for 0.5 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.
	Fig9: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-DDB1 antibody (ET1706-22) at 1/50 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 (ET1706-22) at 1/50 dilution for 0.5 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”.