HA720007

MED4 Recombinant Rabbit Monoclonal Antibody [JE56-55]

cat.: HA720007

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	JE56-55

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 30 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human MED4 aa 100-250.
Positive control:	THP-1 cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, RAW264.7, human small intestine tissue, human brain tissue, human testis tissue, mouse brain tissue, rat brain tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P	1:2,000 1:100 1:200-1:1,000
Uniprot #:	SwissProt: Q9NPJ6 Human \| Q9CQA5 Mouse \| Q561Q8 Rat
Alternative names:	Activator-recruited cofactor 36 kDa component ARC36 DRIP36 FLJ10956 HSPC126 med4 MED4_HUMAN Mediator complex subunit 4 Mediator of RNA polymerase II transcription subunit 4 Mediator of RNA polymerase II transcription, subunit 4 homolog OTTHUMP00000018390 OTTHUMP00000018391 OTTHUMP00000018392 RP11-90M2.2 TRAP/SMCC/PC2 subunit p36 subunit TRAP36 VDRIP Vitamin D3 receptor-interacting protein complex 36 kDa component

Images

Fig1: Western blot analysis of MED4 on different lysates with Rabbit anti-MED4 antibody (HA720007) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: HepG2 cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: Mouse testis tissue lysate
Lane 7: Rat testis tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 30 kDa
Observed band size: 35 kDa

Exposure time: 59 seconds; ECL: K1801;

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 (HA720007) 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.

Fig2: Immunocytochemistry analysis of RAW264.7 cells labeling MED4 with Rabbit anti-MED4 antibody (HA720007) 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-MED4 antibody (HA720007) 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: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-MED4 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA720007, 1/200) for 30 minutes 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-MED4 antibody (HA720007) 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 (HA720007) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-MED4 antibody (HA720007) 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 (HA720007) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-MED4 antibody (HA720007) 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 (HA720007) 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.

Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-MED4 antibody (HA720007) 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 (HA720007) 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”.