HA601038

Ephrin A1 Mouse Monoclonal Antibody [A7B10]

cat.: HA601038

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	A7B10

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 24 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within Human Ephrin A1 aa 13-190 / 205.
Positive control:	HUVEC cell lysates, HUVEC, SK-Br-3, human breast cancer tissue, mouse brain tissue, rat brain tissue.
Subcellular location:	Cell membrane, Secreted.
Recommended Dilutions: WB IF-Cell IHC-P	1:2,000 1:100 1:500
Uniprot #:	SwissProt: P20827 Human \| P52793 Mouse \| P97553 Rat
Alternative names:	B61 ECKLG EFL 1 EFL1 EFNA 1 Efna1 EFNA1_HUMAN EPH related receptor tyrosine kinase ligand 1 EPH-related receptor tyrosine kinase ligand 1 Ephrin-A1 Ephrin-A1, secreted form EphrinA1 EPLG 1 EPLG1 Immediate early response protein B61 LERK 1 LERK-1 LERK1 Ligand of eph related kinase 1 OTTHUMP00000033242 OTTHUMP00000033271 secreted form TNF alpha-induced protein 4 TNFAIP 4 TNFAIP4 Tumor necrosis factor alpha induced protein 4 Tumor necrosis factor alpha-induced protein 4

Images

Fig1: Western blot analysis of Ephrin A1 on HUVEC cell lysates with Mouse anti-Ephrin A1 antibody (HA601038) at 1/2,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 24 kDa
Observed band size: 25 kDa

Exposure time: 3 minutes;

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 (HA601038) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HUVEC cells labeling Ephrin A1 with Mouse anti-Ephrin A1 antibody (HA601038) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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 Mouse anti-Ephrin A1 antibody (HA601038) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunocytochemistry analysis of SK-Br-3 cells labeling Ephrin A1 with Mouse anti-Ephrin A1 antibody (HA601038) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Ephrin A1 antibody (HA601038) at 1/100 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Mouse anti-Ephrin A1 antibody (HA601038) at 1/500 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 (HA601038) at 1/500 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 mouse brain tissue with Mouse anti-Ephrin A1 antibody (HA601038) at 1/500 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 (HA601038) at 1/500 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 brain tissue with Mouse anti-Ephrin A1 antibody (HA601038) at 1/500 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 (HA601038) at 1/500 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”.