ER1803-70

Annexin A1 Rabbit Polyclonal Antibody

cat.: ER1803-70

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 39 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human Annexin A1 aa 150-310.
Positive control:	A431 cell lysate, K-562 cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, A431, NIH/3T3, C6, human esophagus tissue, human placenta tissue, mouse esophagus tissue, rat esophagus tissue.
Subcellular location:	Cell membrane, Cell projection, Cilium, Cytoplasm, Cytoplasmic vesicle, Endosome, Membrane, Nucleus, Secreted.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000-1:2,000 1:100-1:250 1:200-1:1,000 1:1,000
Uniprot #:	SwissProt: P04083 Human \| P10107 Mouse \| P07150 Rat
Alternative names:	Annexin 1 Annexin A1 Annexin I (lipocortin I) Annexin I Annexin-1 AnnexinA1 AnnexinI ANX 1 ANX A1 ANX1 ANXA 1 ANXA1 ANXA1 protein ANXA1_HUMAN Calpactin 2 Calpactin II Calpactin-2 CalpactinII Chromobindin 9 Chromobindin-9 Chromobindin9 HGNC:533 Lipocortin 1 Lipocortin I Lipocortin1 LipocortinI LPC 1 LPC1 p35 Phospholipase A2 inhibitory protein

Images

Fig1: Western blot analysis of Annexin A1 on different lysates with Rabbit anti-Annexin A1 antibody (ER1803-70) at 1/1,000 dilution.

Lane 1: A431 cell lysate
Lane 2: K-562 cell lysate
Lane 3: C2C12 cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 39 kDa
Observed band size: 33/37 kDa

Exposure time: 2 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 (ER1803-70) 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 A431 cells labeling Annexin A1 with Rabbit anti-Annexin A1 antibody (ER1803-70) at 1/250 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 Rabbit anti-Annexin A1 antibody (ER1803-70) at 1/250 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 (M1305-2, 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 Annexin A1 with Rabbit anti-Annexin A1 antibody (ER1803-70) 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 Rabbit anti-Annexin A1 antibody (ER1803-70) 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 (M1305-2, 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 Annexin A1 with Rabbit anti-Annexin A1 antibody (ER1803-70) 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 Rabbit anti-Annexin A1 antibody (ER1803-70) 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 (M1305-2, 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 human esophagus tissue with Rabbit anti-Annexin A1 antibody (ER1803-70) 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 (ER1803-70) 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 human placenta tissue with Rabbit anti-Annexin A1 antibody (ER1803-70) 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 (ER1803-70) 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 mouse esophagus tissue with Rabbit anti-Annexin A1 antibody (ER1803-70) 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 (ER1803-70) 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.

Fig8: Immunohistochemical analysis of paraffin-embedded rat esophagus tissue with Rabbit anti-Annexin A1 antibody (ER1803-70) 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 (ER1803-70) 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.

Fig9: Flow cytometric analysis of A431 cells labeling Annexin A1.

Cells were fixed and permeabilized. Then stained with the primary antibody (ER1803-70, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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