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Rabbit Recombinant Monoclonal HIF-1 alpha antibody. Carrier free. Suitable for IP, ChIC/CUT&RUN-seq, ChIP-seq, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 1 publication.


Images

Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (AB221610), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (AB221610), expandable thumbnail
  • Immunoprecipitation - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (AB221610), expandable thumbnail
  • ChIC/CUT&RUN sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (AB221610), expandable thumbnail
  • ChIP-sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (AB221610), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPChIC/CUT&RUN-seqChIP-seqWBICC/IF
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Expected
Tested
Expected
Rat
Expected
Expected
Expected
Tested
Expected
Cow
Predicted
Predicted
Predicted
Predicted
Predicted
Ferret
Predicted
Predicted
Predicted
Predicted
Predicted
Primates
Predicted
Predicted
Predicted
Predicted
Predicted
Rabbit
Predicted
Predicted
Predicted
Predicted
Predicted
Sheep
Predicted
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Sheep, Cow, Ferret, Primates, Rabbit

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Sheep, Cow, Ferret, Primates, Rabbit

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Sheep, Cow, Ferret, Primates, Rabbit

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

-

Notes

HIF-1 alpha expression is absent in most normal tissues (PMID: 12128120, 24835245, 11689469, 20217131). The antibody only works in hypoxic cell and tissue lysates.

Species

Rat

Dilution info

-

Notes

HIF-1 alpha expression is absent in most normal tissues (PMID: 12128120, 24835245, 11689469, 20217131). The antibody only works in hypoxic cell and tissue lysates.

Species

Human

Dilution info

-

Notes

HIF-1 alpha expression is absent in most normal tissues (PMID: 12128120, 24835245, 11689469, 20217131). The antibody only works in hypoxic cell and tissue lysates.

Predicted
Predicted

Species

Sheep, Cow, Ferret, Primates, Rabbit

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Sheep, Cow, Ferret, Primates, Rabbit

Dilution info

-

Notes

-

Associated Products

Select an associated product type

12 products for Alternative Product

2 products for Alternative Version

Target data

Function

Functions as a master transcriptional regulator of the adaptive response to hypoxia (PubMed:11292861, PubMed:11566883, PubMed:15465032, PubMed:16973622, PubMed:17610843, PubMed:18658046, PubMed:20624928, PubMed:22009797, PubMed:30125331, PubMed:9887100). Under hypoxic conditions, activates the transcription of over 40 genes, including erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, HILPDA, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia (PubMed:11292861, PubMed:11566883, PubMed:15465032, PubMed:16973622, PubMed:17610843, PubMed:20624928, PubMed:22009797, PubMed:30125331, PubMed:9887100). Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease (PubMed:22009797). Heterodimerizes with ARNT; heterodimer binds to core DNA sequence 5'-TACGTG-3' within the hypoxia response element (HRE) of target gene promoters (By similarity). Activation requires recruitment of transcriptional coactivators such as CREBBP and EP300 (PubMed:16543236, PubMed:9887100). Activity is enhanced by interaction with NCOA1 and/or NCOA2 (PubMed:10594042). Interaction with redox regulatory protein APEX1 seems to activate CTAD and potentiates activation by NCOA1 and CREBBP (PubMed:10202154, PubMed:10594042). Involved in the axonal distribution and transport of mitochondria in neurons during hypoxia (PubMed:19528298).(Microbial infection) Upon infection by human coronavirus SARS-CoV-2, is required for induction of glycolysis in monocytes and the consequent pro-inflammatory state (PubMed:32697943). In monocytes, induces expression of ACE2 and cytokines such as IL1B, TNF, IL6, and interferons (PubMed:32697943). Promotes human coronavirus SARS-CoV-2 replication and monocyte inflammatory response (PubMed:32697943).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal HIF-1 alpha antibody. Carrier free. Suitable for IP, ChIC/CUT&RUN-seq, ChIP-seq, WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EPR16897

Purification technique

Affinity purification Protein A

Specificity

HIF-1 alpha expression is absent in most normal tissues (PMID: 12128120, 24835245, 11689469, 20217131). The antibody only works in hypoxic cell and tissue lysates.

FURTHER INFORMATION ON SPECIFICITY(Chinese Version) available under the support & downloads section.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab221610 is the carrier-free version of Anti-HIF-1 alpha antibody [EPR16897] ab179483.

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

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

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.

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.

Supplementary info

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

Activity summary

HIF-1 alpha also known as hypoxia-inducible factor 1-alpha is a transcription factor critical in cellular response to low oxygen levels. Its molecular weight usually ranges from 93 to 120 kDa. You can find HIF-1 alpha expressed in tissues throughout the body but its expression significantly increases under hypoxic conditions. Researchers often use the HIF-1a ELISA to measure its expression levels. HIF-1 alpha forms a complex with other proteins to perform its functions effectively.

Biological function summary

HIF-1 alpha regulates gene expression in response to hypoxic conditions in cells. It forms a complex with HIF-1 beta to activate transcription of various genes involved in energy metabolism angiogenesis and erythropoiesis. HIF-1 alpha enables cells to adapt to reduced oxygen availability allowing for cellular survival and function under stress. It plays an important role in promoting the expression of genes like VEGF and EPO which are important for vascular and red blood cell development respectively.

Pathways

HIF-1 alpha plays an integral role in the hypoxia signaling pathway and the glycolytic pathway. In the hypoxia signaling pathway HIF-1 alpha partners with VHL (Von Hippel-Lindau) protein that regulates its degradation under normal oxygen conditions. When oxygen levels drop HIF-1 alpha avoids degradation stabilizes and translocates into the nucleus to initiate transcription of hypoxia-responsive genes. The glycolytic pathway involvement highlights its function in adapting energy production under hypoxic conditions through collaboration with enzymes and transporters associated with glycolysis.

Associated diseases and disorders

HIF-1 alpha has been implicated in cancer and ischemic diseases. Its role in promoting angiogenesis and metabolic adaptation makes it a contributor to tumor growth and survival collaborating with oncogenes such as c-Myc. In ischemic diseases like stroke or myocardial infarction HIF-1 alpha's ability to induce protective responses can mitigate tissue damage through regulation of survival pathways. Understanding these interactions helps in the development of therapeutic strategies targeting HIF-1 alpha in disease contexts.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Lanes 1 - 6: Merged signal (red and green). Green - Anti-HIF-1 alpha antibody [EPR16897] ab179483 observed at 105 kDa. Red - loading control, Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade ab24834, observed at 50 kDa.

    Anti-HIF-1 alpha antibody [EPR16897] ab179483 was shown to specifically react with HIF-1 alpha in wild-type HAP1 treated DMOG (0.5mM 18hr) cells as signal was lost in HAP1 knockout treated DMOG (0.5mM 18hr) knockout cells. Wild-type and HAP1 knockout samples were subjected to SDS-PAGE. Anti-HIF-1 alpha antibody [EPR16897] ab179483 and Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade ab24834 (Mouse anti-Histone H3 loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

    All lanes: Western blot - Anti-HIF-1 alpha antibody [EPR16897] (Anti-HIF-1 alpha antibody [EPR16897] ab179483) at 1/1000 dilution

    Lane 1: Wild-type HAP1 whole cell lysate at 40 µg

    Lane 2: Wild type HAP1 treated with DMOG (0.5mM 18hr) whole cell lysate at 40 µg

    Lane 3: HIF1A knockout HAP1 whole cell lysate at 40 µg

    Lane 4: HIF1A knockout HAP1 treated with DMOG (0.5mM 18hr) whole cell lysate at 40 µg

    Lane 5: HeLa whole cell lysate at 40 µg

    Lane 6: HeLa treated with DMOG (0.5mM 18hr) whole cell lysate at 40 µg

    Predicted band size: 92 kDa

    Observed band size: 105 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling HIF-1-alpha with Anti-HIF-1 alpha antibody [EPR16897] ab179483 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cells treated with DFO (1 mM, 24 h). The nuclear counter stain is DAPI (blue). Tubulin is detected with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 (anti-alpha Tubulin mouse mAb) (Alexa Fluor® 594) at 1/200 dilution (red).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

  • Immunoprecipitation - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    Immunoprecipitation - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Anti-HIF-1 alpha antibody [EPR16897] ab179483 at 1/30 dilution immunoprecipitating HIF-1 alpha in HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate. Western blot was performed from the immunoprecipitate using Anti-HIF-1 alpha antibody [EPR16897] ab179483 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate 10 µg
    Lane 2: Anti-HIF-1 alpha antibody [EPR16897] ab179483 IP in HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HIF-1 alpha antibody [EPR16897] ab179483 in HeLa treated with 500 µM CoCl2 for 24 hours whole cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 8 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

    Lane 2: Immunoprecipitation - Anti-HIF-1 alpha antibody [EPR16897] (Anti-HIF-1 alpha antibody [EPR16897] ab179483) at 1/30 dilution

    Lane 2: Immunoprecipitation - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    All lanes: HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate at 10 µg

    Secondary

    All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Observed band size: 110 kDa

    Exposure time: 8s

    Anti-HIF-1 alpha antibody [EPR16897] ab179483 at 1/30 dilution immunoprecipitating HIF-1 alpha in HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate. Western blot was performed from the immunoprecipitate using Anti-HIF-1 alpha antibody [EPR16897] ab179483 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.

    Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate 10 µg
    Lane 2: Anti-HIF-1 alpha antibody [EPR16897] ab179483 IP in HeLa (human cervix adenocarcinoma epithelial cell) treated with 500 µM CoCl2 for 24 hours whole cell lysate
    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HIF-1 alpha antibody [EPR16897] ab179483 in HeLa treated with 500 µM CoCl2 for 24 hours whole cell lysate


    Blocking/Dilution buffer: 5% NFDM/TBST.
    Exposure time: 8 seconds.

  • ChIC/CUT&RUN sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    ChIC/CUT&RUN sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells treated with Cocl2 (500 μM 20h+4h) and MG-132 (10µM 4h) and 5 µg of Anti-HIF-1 alpha antibody [EPR16897] ab179483 [EPR16897]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 is also shown.

    Additional screenshots of mapped reads can be found in the Protocol booklet in the Support and downloads section.
    The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

  • ChIP-sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    ChIP-sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Chromatin was prepared from HeLa cells treated with CoCl2 (350μM 20h+4h) add MG-132(10µM 4h). Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 8 µg of Anti-HIF-1 alpha antibody [EPR16897-145] ab308433 [EPR16897-145] or Anti-HIF-1 alpha antibody [EPR16897] ab179483 [EPR16897]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

  • Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST
    Lysates were freshly made and used immediately to minimise protein degradation.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

    Lanes 1 - 4: Western blot - Anti-HIF-1 alpha antibody [EPR16897] (Anti-HIF-1 alpha antibody [EPR16897] ab179483) at 1/1000 dilution

    Lanes 1 - 4: Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Lane 1: Untreated HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HT-1080 treated with 0.5 mM DFO for 24 hours whole cell lysate at 20 µg

    Lane 3: HT-1080 treated with 0.5 mM DFO for 24 hours, then added 10uM MG-132 for 4 hours, whole cell lysate at 20 µg

    Lane 4: HT-1080 treated with 10uM MG-132 for 4 hours, whole cell lysate at 20 µg

    Observed band size: 50-110 kDa

    Exposure time: 10s

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST
    Lysates were freshly made and used immediately to minimise protein degradation.

  • ChIP-sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    ChIP-sequencing - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    Chromatin was prepared from HeLa cells treated with CoCl2 (350μM 20h+4h) add MG-132(10µM 4h). Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 107 cells and 8 µg of Anti-HIF-1 alpha antibody [EPR16897-145] ab308433 [EPR16897-145] or Anti-HIF-1 alpha antibody [EPR16897] ab179483 [EPR16897]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

  • Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610), expandable thumbnail

    Western blot - Anti-HIF-1 alpha antibody [EPR16897] - BSA and Azide free (ab221610)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HIF-1 alpha antibody [EPR16897] ab179483).

    HIF-1-alpha is constantly synthesized but rapidly degraded under normoxic conditions, whereas reduced oxygen concentration results in stabilization of HIF-1-alpha. (PMID: 15104534)

    Exposure time: Lanes 1-4:10 seconds, Lanes 5-6: 20 seconds

    All lanes: Western blot - Anti-HIF-1 alpha antibody [EPR16897] (Anti-HIF-1 alpha antibody [EPR16897] ab179483) at 1/1000 dilution

    Lanes 1 and 3: Untreated HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

    Lane 2: HeLa treated with 0.5mM CoCl2 for 6h whole cell lysate at 20 µg

    Lane 4: HeLa treated with 0.5mM DFO for 24h whole cell lysate at 20 µg

    Lane 5: Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

    Lane 6: NIH/3T3 (Mouse embryonic fibroblast) treated with 0.1mM CoCl2 for 48h whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 50-110 kDa

Downloads

Product protocols

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

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

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