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[KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from wild type (WT) and Annexin A1/ANXA1 knockdown (KD) HeLa cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at 1:7900 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 0.5s.

Western blot analysis of lysates from C2C12 cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at 1:7900 dilution incubated overnight at 4℃.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 0.5s.

ABclonal:Immunohistochemistry - [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918)

Immunohistochemistry analysis of paraffin-embedded Human liver cancer tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Human tonsil tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:9000 (40x lens). High pressure antigen retrieval performed with 0.01M Tris-EDTA Buffer(pH 9.0) prior to IHC staining.

ABclonal:Immunofluorescence - [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918)

Confocal imaging of HeLa cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of C2C12 cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

Confocal imaging of U-251 MG cells using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). The cells were counterstained with α-Tubulin Mouse mAb (AC012, dilution 1:400) followed by incubation with ABflo® 488-conjugated Goat Anti-Mouse IgG (H+L) Ab (AS076, dilution 1:500) (Green). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918)

Immunoprecipitation of [KD Validated] Annexin A1/ANXA1 from 500 µg extracts of C2C12 cells was performed using 2 µg of [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using [KD Validated] Annexin A1/ANXA1 Rabbit mAb (A25918) at a dilution of 1:8000.

Overview

Product name	[KD Validated] Annexin A1/ANXA1 Rabbit mAb
Catalog No.	A25918
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC66904

Background

This gene encodes a membrane-localized protein that binds phospholipids. This protein inhibits phospholipase A2 and has anti-inflammatory activity. Loss of function or expression of this gene has been detected in multiple tumors.

Immunogen information

Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 2-346 of human Annexin A1/ANXA1 (NP_000691.1).
Sequence	AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALHKAIMVKGVDEATIIDILTKRNNAQRQQIKAAYLQETGKPLDETLKKALTGHLEEVVLALLKTPAQFDADELRAAMKGLGTDEDTLIEILASRTNKEIRDINRVYREELKRDLAKDITSDTSGDFRNALLSLAKGDRSEDFGVNEDLADSDARALYEAGERRKGTDVNVFNTILTTRSYPQLRRVFQKYTKYSKHDMNKVLDLELKGDIEKCLTAIVKCATSKPAFFAEKLHQAMKGVGTRHKALIRIMVSRSEIDMNDIKAFYQKMYGISLCQAILDETKGDYEKILVALCGGN
Gene ID
Swiss Prot
Synonyms	ANX1; LPC1
Calculated MW	39kDa
Observed MW	38kDa/

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:7000 - 1:70000 IHC-P 1:2000 - 1:20000 IF/ICC 1:200 - 1:400 IP 0.5μg-4μg antibody for 400μg-600μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% Sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence Immunoprecipitation
Positive samples	HeLa, C2C12, C2C12
Cellular location	Apical cell membrane, Basolateral cell membrane, Cell membrane, Cell projection, Cytoplasm, Cytoplasmic vesicle, Cytoplasmic vesicle membrane, Early endosome, Endosome membrane, Extracellular side, Lateral cell membrane, Membrane, Nucleus, Peripheral membrane protein, Secreted, cilium, exosome, extracellular space, phagocytic cup, secretory vesicle lumen.