AB316759

Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free

Recombinant
RabMAb
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Rabbit Recombinant Monoclonal ErbB2 / HER2 phospho Y1221 + Y1222 antibody. Carrier free. Suitable for Dot, ICC/IF, WB and reacts with Synthetic peptide - Human, Human, Mouse samples.

View Alternative Names

CD340, HER2, MLN19, NEU, NGL, ERBB2, Receptor tyrosine-protein kinase erbB-2, Metastatic lymph node gene 19 protein, Proto-oncogene Neu, Proto-oncogene c-ErbB-2, Tyrosine kinase-type cell surface receptor HER2, p185erbB2, MLN 19

8 Images

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling ErbB2 / HER2 (phospho Y1221 + Y1222) with ab316758 at 1/100 (5.14 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing increased membranous and cytoplasmic staining in HeLa cells starved for 4 hours then treated with EGF (200 ng/ml) for 15 min. The signal decreased after alkaline phosphatase treatment at 37℃ for 2h. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-BR-3 (human breast adenocarcinoma epithelial cell) cells labelling ErbB2 / HER2 (phospho Y1221 + Y1222) with ab316758 at 1/100 (5.14 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing mainly membranous staining in SK-BR-3 cells, then the signal decreased after phosphatase treatment at 37℃ for 2h. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 4T1 (mouse mammary gland carcinoma epithelial cell) cells labelling ErbB2 / HER2 (phospho Y1221 + Y1222) with ab316758 at 1/100 (5.14 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing membranous and cytoplasmic staining in 4T1 cells, and the signal decreased after phosphatase treatment at 37℃ for 2h. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Supplier Data

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

ErbB2 / HER2 (phospho Y1221 + Y1222) expression can be downregulated by lapatinib treatment (PMID : 27738326).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-ErbB2 / HER2 antibody - Total protein control (ab214275) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] (<a href='/en-us/products/primary-antibodies/erbb2-her2-phospho-y1221-y1222-antibody-epr28391-67-ab316758'>ab316758</a>) at 1/1000 dilution

Lane 1:

Untreated SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

SK-BR-3 treated with 5uM lapatinib for 24 hours whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,36 kDa

false

Exposure time: 180s

Supplier Data

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-ErbB2 / HER2 antibody - Total protein control (ab214275) staining at 1/1000 dilution.

All lanes:

Lane 1:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,36 kDa

false

Exposure time: 92s

Supplier Data

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

ErbB2 / HER2 (phospho Y1221 + Y1222) expression can be induced by EGF treatment (PMID : 7728765).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-ErbB2 / HER2 antibody - Total protein control (ab214275) staining at 1/1000 dilution.

All lanes:

Lane 1:

Untreated HeLa (human cervix adenocarcinoma epithelial cell) starved for 4 hours whole cell lysate (untreated membrane) at 20 µg

Lane 2:

HeLa starved for 4 hours, then treated with /ml EGF for 15 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

HeLa starved for 4 hours, then treated with /ml EGF for 15 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,15 kDa

false

Exposure time: 81s

Supplier Data

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) staining at 1/100000 dilution.

In Western blot, Anti-ErbB2 / HER2 antibody - Total protein control (ab214275) staining at 1/1000 dilution.

All lanes:

Lane 1:

Untreated T47D (human ductal breast epithelial tumor epithelial cell) starved for 4 hours whole cell lysate (untreated membrane) at 20 µg

Lane 2:

T47D starved for 4 hours, then treated with /ml Neuregulin for 10 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

T47D starved for 4 hours, then treated with /ml Neuregulin for 10 minutes whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 180 kDa,15 kDa

false

Exposure time: 103s

Supplier Data

Dot Blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

This data was developed using ab316758, the same antibody clone in a different buffer formulation.

Dot blot analysis of ErbB2 / HER2 (phospho Y1221 + Y1222) using ab316758 at 1 : 1000 (0.514 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lane 1 : ErbB2 / HER2 (phospho Y1221 + Y1222) peptide a
Lane 2 : ErbB2 / HER2 (phospho Y1221 + Y1222) peptide b
Lane 3 : ErbB2 / HER2 (phospho Y1221 ) peptide c
Lane 4 : ErbB2 / HER2 (phospho Y1222) peptide d
Lane 5 : ErbB2 / HER2 non-phospho peptide e

The antibody recognizes ERBB2 protein when phosphorylated at Tyr1221 and Tyr1222, and shows slight reactivity with ERBB2 when singly phosphorylated at Tyr1221 or Tyr1222.

All lanes:

Dot Blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] (<a href='/en-us/products/primary-antibodies/erbb2-her2-phospho-y1221-y1222-antibody-epr28391-67-ab316758'>ab316758</a>) at 1/1000 dilution

Lane 1:

ErbB2 / HER2 (phospho Y1221 + Y1222) peptide a

Lane 2:

ErbB2 / HER2 (phospho Y1221 + Y1222) peptide b

Lane 3:

ErbB2 / HER2 (phospho Y1221 ) peptide c

Lane 4:

ErbB2 / HER2 (phospho Y1222) peptide d

Lane 5:

ErbB2 / HER2 non-phospho peptide e

Secondary

All lanes:

Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 180s

Unconjugated

Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67]

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR28391-67

Isotype

IgG

Carrier free

Yes

Reacts with

Human, Mouse

Applications

Dot, ICC/IF, WB

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

Unsuitable for mouse WB.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "Dot" : {"fullname" : "Dot Blot", "shortname":"Dot"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IHCFr" : {"fullname" : "Immunohistochemistry (Frozen sections)", "shortname":"IHC-Fr"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Mouse": { "Dot-species-checked": "guaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Rat": { "Dot-species-checked": "notRecommended", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" }, "Synthetic peptide - Human": { "Dot-species-checked": "testedAndGuaranteed", "Dot-species-dilution-info": "", "Dot-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "WB-species-checked": "notRecommended", "WB-species-dilution-info": "", "WB-species-notes": "", "IHCP-species-checked": "notRecommended", "IHCP-species-dilution-info": "", "IHCP-species-notes": "", "IHCFr-species-checked": "notRecommended", "IHCFr-species-dilution-info": "", "IHCFr-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "" } } }

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Product details

ab316759 is the carrirer-free version of ab316758.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

The ErbB2 also known as HER2 or HER2 protein is a transmembrane receptor protein with a molecular weight of about 185 kDa. It serves as part of the epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases. The ErbB2 protein is expressed mainly in epithelial tissues including those of the breast and the gastrointestinal tract. It lacks a known ligand-binding domain which sets it apart from other members of its family. Due to its structure ErbB2 dimerizes with other members of the EGFR family to exert its effects in cellular signaling.

Biological function summary

The role of ErbB2 extends beyond a singular function. It becomes an active component when forming heterodimers with other EGFR family members such as ErbB3 to initiate various downstream signaling cascades. The dimerization activates intracellular pathways leading to cell proliferation survival differentiation and migration. Within cells ErbB2 influences processes critical for normal development and tissue homeostasis contributing significantly to signal transduction networks.

Pathways

ErbB2 plays a pivotal role in pathways such as the PI3K/Akt and MAPK/ERK pathways. These pathways are important in mediating cellular responses to growth signals. The PI3K/Akt pathway activated by HER2 signaling regulates cell growth and survival while the MAPK/ERK pathway contributes to cell differentiation and proliferation. ErbB2's interaction with proteins like ErbB3 is fundamental in these pathways increasing the amplitude and diversity of downstream signals.

ErbB2 is significantly associated with breast cancer and gastric cancer. The overexpression or gene amplification of HER2 is observed in approximately 20% of breast cancer cases correlating with aggressive tumor growth and poor prognosis. ErbB2-related signaling contributes to oncogenic processes by promoting excessive cell proliferation. Targeting ErbB2 in these cancers is common using therapies such as monoclonal antibodies like trastuzumab. In the context of gastric cancer the role of ErbB2 mirrors its function in breast cancer and targeting HER2 holds therapeutic potential.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

Protein tyrosine kinase that is part of several cell surface receptor complexes, but that apparently needs a coreceptor for ligand binding. Essential component of a neuregulin-receptor complex, although neuregulins do not interact with it alone. GP30 is a potential ligand for this receptor. Regulates outgrowth and stabilization of peripheral microtubules (MTs). Upon ERBB2 activation, the MEMO1-RHOA-DIAPH1 signaling pathway elicits the phosphorylation and thus the inhibition of GSK3B at cell membrane. This prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization.. In the nucleus is involved in transcriptional regulation. Associates with the 5'-TCAAATTC-3' sequence in the PTGS2/COX-2 promoter and activates its transcription. Implicated in transcriptional activation of CDKN1A; the function involves STAT3 and SRC. Involved in the transcription of rRNA genes by RNA Pol I and enhances protein synthesis and cell growth.

See full target information ERBB2 pY1221 + Y1222

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Western blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Dot Blot - Anti-ErbB2 / HER2 (phospho Y1221 + Y1222) antibody [EPR28391-67] - BSA and Azide free (AB316759)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Specificity

Complementary Products

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Product promise